Self-assembled peptide-substance P hydrogels alleviate inflammation and ameliorate the cartilage regeneration in knee osteoarthritis.

BACKGROUND: Self-assembled peptide (SAP)-substance P (SP) hydrogels can be retained in the joint cavity longer than SP alone, and they can alleviate inflammation and ameliorate cartilage regeneration in knee osteoarthritis (OA). We conducted a preclinical study using diverse animal models of OA and an in vitro study using human synoviocytes and patient-derived synovial fluids to demonstrate the effect of SAP-SP complex on the inflammation and cartilage regeneration. METHODS: Surgical induction OA model was prepared with New Zealand white female rabbits and chemical induction, and naturally occurring OA models were prepared using Dunkin Hartely female guinea pigs. The SAP-SP complex or control (SAP, SP, or saline) was injected into the joint cavities in each model. We performed micro-computed tomography (Micro-CT) analysis, histological evaluation, immunofluorescent analysis, and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling (TUNEL) assay and analyzed the recruitment of intrinsic mesenchymal stem cells (MSCs), macrophage activity, and inflammatory cytokine in each OA model. Human synoviocytes were cultured in synovial fluid extracted from human OA knee joints injected with SAP-SP complexes or other controls. Proliferative capacity and inflammatory cytokine levels were analyzed. RESULTS: Alleviation of inflammation, inhibition of apoptosis, and enhancement of intrinsic MSCs have been established in the SAP-SP group in diverse animal models. Furthermore, the inflammatory effects on human samples were examined in synoviocytes and synovial fluid from patients with OA. In this study, we observed that SAP-SP showed anti-inflammatory action in OA conditions and increased cartilage regeneration by recruiting intrinsic MSCs, inhibiting progression of OA. CONCLUSIONS: These therapeutic effects have been validated in diverse OA models, including rabbits, Dunkin Hartley guinea pigs, and human synoviocytes. Therefore, we propose that SAP-SP may be an effective injectable therapeutic agent for treating OA. In this manuscript, we report a preclinical study of novel self-assembled peptide (SAP)-substance P (SP) hydrogels with diverse animal models and human synoviocytes and it displays anti-inflammatory effects, apoptosis inhibition, intrinsic mesenchymal stem cells recruitments and cartilage regeneration.

Nanotopographic cues and stiffness control of tendon-derived stem cells from diverse conditions.

BACKGROUND: Tendon-derived stem cells (TDSCs) are key factors associated with regeneration and healing in tendinopathy. The aim of this study was to investigate the effects of mechanical stiffness and topographic signals on the differentiation of TDSCs depending on age and pathological conditions. MATERIALS AND METHODS: We compared TDSCs extracted from normal tendon tissues with TDSCs from tendinopathic Achilles tendon tissues of Sprague Dawley rats in vitro and TDSCs cultured on nanotopographic cues and substrate stiffness to determine how to control the TDSCs. The tendinopathy model was created using a chemical induction method, and the tendon injury model was created via an injury-and-overuse method. Norland Optical Adhesive 86 (NOA86) substrate with 2.48 GPa stiffness with and without 800 nm-wide nanogrooves and a polyurethane substrate with 800 nm-wide nanogrooves were used. RESULTS: TDSCs from 5-week-old normal tendon showed high expression of type III collagen on the flat NOA86 substrate. In the 15-week normal tendon model, expression of type III collagen was high in TDSCs cultured on the 800 nm NOA86 substrates. However, in the 15-week tendon injury model, expression of type III collagen was similar irrespective of nanotopographic cues or substrate stiffness. The expression of type I collagen was also independent of nanotopographic cues and substrate stiffness in the 15-week normal and tendon injury models. Gene expression of scleraxis was increased in TDSCs cultured on the flat NOA86 substrate in the 5-week normal tendon model (P=0.001). In the 15-week normal tendon model, scleraxis was highly expressed in TDSCs cultured on the 800 nm and flat NOA86 substrate (P=0.043). However, this gene expression was not significantly different between the substrates in the 5-week tendinopathy and 15-week tendon injury models. CONCLUSION: Development and maturation of tendon are enhanced when TDSCs from normal tendons were cultured on stiff surface, but not when the TDSCs came from pathologic models. Therapeutic applications of TDSCs need to be flexible based on tendon age and tendinopathy.

Development and characterization of various osteoarthritis models for tissue engineering.

Osteoarthritis (OA) is characterized by a progressive loss of articular cartilage, subchondral bone sclerosis and synovial inflammation and is the most common chronic condition worldwide today. However, most treatments have focused on pain relief and OA symptoms. For these reasons, many ongoing studies are currently trying to develop efficient and successful therapies based on its pathology. Animal models that mimic the histopathology and symptoms of OA have a critical role in OA research and make it possible to investigate both secondary osteoarthritic changes due to a precedent event such as traumatic injury and naturally occurring changes for the development of therapeutics which can be tested in preclinical and clinical OA trials. We induced OA in various animal models including rats, rabbits and guinea pigs by chemical, surgical and naturally occurring methods. In particular, the Dunkin-Hartley guinea pig is very attractive as an OA animal model because OA slowly progresses which is similar to human primary OA. Thus, this animal model mimics the pathophysiological process and environment of human primary OA. Besides the spontaneous OA model, anterior cruciate ligament transection (ACLT) with medial meniscectomy and bilateral ovariectomy (OVX) as well as a chemical technique using sodium monoiodoacetate (MIA) were used to induce OA. We found that ACLT in the rat model induced OA changes in the histology and micro-CT image compared to OVX. The osteoarthritic change significantly increased following ACLT surgery in the rabbit model. Furthermore, we identified that OA pathogenic changes occurred in a time-dependent manner in spontaneous Dunkin-Hartley guinea pigs. The MIA injection model is a rapid and minimally invasive method for inducing OA in animal models, whereas the spontaneous OA model has a slow and gradual progression of OA similar to human primary OA. We observed that histological OA change was extraordinarily increased at 9 ½ months in the spontaneous OA model, and thus, the grade was similar with that of the MIA model. Therefore, this study reports on OA pathology using various animal models as well as the spontaneous results naturally occurring in an OA animal model which had developed cartilage lesions and progressive osteoarthritic changes.

Effect of high frequency electromagnetic wave stimulation on muscle injury in a rat model.

INTRODUCTION: The aim of this study was to investigate biological changes in tissues with muscle contusion after the application of high frequency (HF) electromagnetic wave. METHODS: An acrylic pipe was placed on the right hind limb and a metallic ball was dropped inside the pipe, which resulted in a muscle contusion. After acquiring the optimal condition for muscle contusion, 20 Sprague-Dawley rats were allocated to the HF treatment (N = 10) and sham groups (N = 10), which then underwent muscle contusion injury at their right thigh. The thickness and circumference of the right thigh and the left thigh (negative control groups) were measured (day 0). HF electromagnetic wave stimulation for three days was performed on the contusion area in the HF group after one day. Thickness was measured at the thickest area of both hind limbs and the circumference was measured every day for three days. The sham group received no treatment, and the circumference and thickness were measured using the same method. After three days, Hematoxylin and eosin and immunohistochemical (IHC) staining for IL-1ß were performed and TUNEL assay was conducted for apoptosis in the skin and muscle layers. RESULTS: The thigh muscle thickness at day 1 was significantly different between groups (P = 0.018) and this difference was observed between both sham and control groups (corrected P = 0.007), and between sham and HF groups (corrected P = 0.043). Thigh circumference was significantly different at day 3 (P = 0.047) and this difference was found between sham and control groups (corrected P = 0.018), and between sham and HF groups (corrected P = 0.032). In the HF group, the inflammatory response was reduced to almost the same level as the control group. Evaluation of IL-1ß level, the inflammatory cytokine, through IHC showed marked localization of IL-1ß in muscle fibers of the sham group. However, significantly less IL-1ß was observed in the muscle of the HF treatment group. There was neither injury nor apoptosis after HF stimulation. CONCLUSIONS: Application of the HF showed therapeutic effect on muscle contusion by reducing muscle swelling. This effect might be caused by the anti-inflammatory action of the HF, which evoked energy into the injured muscle.

Characteristics of tendon derived stem cells according to different factors to induce the tendinopathy.

Tendon derived stem cells (TDSCs) have been used as a therapeutic agent and as a healing marker. However, there has been no study about the characteristics of TDSCs extracted from tendinopathic tendon tissues. The aim of this study was to find the different characteristics of TDSCs according to the factors to induce the tendinopathy. Five- and fifteen-week old Sprague Dawley rats were used for this study and chemically-induced and injury-induced tendinopathy models were made depending on the age of the animal for different types of tendinopathy. TDSCs from chemically-induced tendinopathy showed markedly low proliferation compared to those from age-matched normal control and injury-induced tendinopathy. In addition, TDSCs from chemically-induced tendinopathy progressed to osteogenesis under an osteogenic differentiation environment more than those from other groups. In contrast, TDSCs from injury-induced tendinopathy showed markedly high proliferation and high expression of type III collagen and α-SMA compared to other groups. Adipogenic potentials in TDSCs from injury-induced tendinopathy were also higher. These different characteristics might be helpful in the development new therapeutic agents for tendon regeneration according to different factors to induce the tendinopathy.

Effects of bone marrow aspirate concentrate and platelet-rich plasma on patients with partial tear of the rotator cuff tendon.

BACKGROUND: We compared the clinical course of rotator cuff tears between rotator cuff exercise and bone marrow aspirate concentration (BMAC)-platelet rich plasma (PRP) injection to identify the therapeutic effects of BMAC-PRP on partial tear of the rotator cuff tendon. METHODS: Twenty-four patients with partial tear of the rotator cuff tendon participated in this study. Twelve patients underwent extraction of BMACs and PRP and received the injection of BMAC-PRP at the tear site under ultrasound guidance. Twelve patients in the control group were asked to perform the rotator cuff exercise for 3 months. Visual analog scale (VAS) and manual muscle test (MMT) scores of the supraspinatus muscle were measured, and the American Shoulder and Elbow Surgeons (ASES) score was recorded before, 3 weeks, and 3 months after injection. Tear size was measured by the greatest longitudinal tear length. RESULTS: The change in the VAS differed between groups at 3 months (P = 0.039) but not at 3 weeks (P = 0.147). The ASES scores in the BMAC-PRP group changed from 39.4 ± 13.0 to 54.5 ± 11.5 at 3 weeks and 74.1 ± 8.5 at 3 months while those in the control group changed from 45.9 ± 12.4 to 56.3 ± 12.3 at 3 weeks (P = 0.712) and 62.2 ± 12.2 at 3 months (P = 0.011). The tear size decreased at 3 weeks or 3 months after the BMAC-PRP injection but was not significantly different from that in the control group. CONCLUSIONS: BMAC-PRP improved pain and shoulder function in patients with partial tear of the rotator cuff tendon. TRIAL REGISTRATION: The patients were registered in the institutional board registry of Samsung Medical Center (registry number 2014-07-173 ).

Enhanced Efficiency in Dye-Sensitized Solar Cells by Electron Transport and Light Scattering on Freestanding TiO2 Nanotube Arrays.

Dye-sensitized solar cells (DSSCs) were fabricated with closed- or open-ended freestanding TiO2 nanotube arrays as photoelectrodes that were decorated with carbon materials and large TiO2 nanoparticles (NPs) to enhance energy conversion efficiency. The energy conversion efficiency of DSSCs based on open-ended freestanding TiO2 nanotube arrays increased from 4.47% to 5.39%, compared to the DSSCs based on closed-ended freestanding TiO2 nanotube arrays. In DSSCs based on the open-ended freestanding TiO2 nanotube arrays, the energy conversion efficiency with carbon materials increased from 5.39% to 6.19% due to better electron transport, and that with a scattering layer from 5.39% to 6.24% due to more light harvesting compared to the DSSCs without carbon materials or scattering layer. Moreover, the energy conversion efficiency of DSSCs based on the open-ended freestanding TiO2 nanotube arrays with both carbon materials and scattering layer increased from 5.39% to 6.98%, which is an enhancement of 29.50%. In DSSCs based on the TiO2 nanotube arrays, the carbon materials can improve electron transport by π-π conjugation, and the large TiO2 NPs can enhance the capacity to light-harvest by scattering.

Multi-Shaped Ag Nanoparticles in the Plasmonic Layer of Dye-Sensitized Solar Cells for Increased Power Conversion Efficiency.

The use of dye-sensitized solar cells (DSSCs) is widespread owing to their high power conversion efficiency (PCE) and low cost of manufacturing. We prepared multi-shaped Ag nanoparticles (NPs) and introduced them into DSSCs to further enhance their PCE. The maximum absorption wavelength of the multi-shaped Ag NPs is 420 nm, including the shoulder with a full width at half maximum (FWHM) of 121 nm. This is a broad absorption wavelength compared to spherical Ag NPs, which have a maximum absorption wavelength of 400 nm without the shoulder of 61 nm FWHM. Therefore, when multi-shaped Ag NPs with a broader plasmon-enhanced absorption were coated on a mesoporous TiO2 layer on a layer-by-layer structure in DSSCs, the PCE increased from 8.44% to 10.22%, equivalent to an improvement of 21.09% compared to DSSCs without a plasmonic layer. To confirm the plasmon-enhanced effect on the composite film structure in DSSCs, the PCE of DSSCs based on the composite film structure with multi-shaped Ag NPs increased from 8.58% to 10.34%, equivalent to an improvement of 20.51% compared to DSSCs without a plasmonic layer. This concept can be applied to perovskite solar cells, hybrid solar cells, and other solar cells devices.

Effect of Bone Marrow Aspirate Concentrate-Platelet-Rich Plasma on Tendon-Derived Stem Cells and Rotator Cuff Tendon Tear.

Bone marrow aspirate concentrates (BMACs) and platelet-rich plasma (PRP) are good sources to control the differentiation of tendon-derived stem cells (TDSCs), but there has been no study about the effect of the BMAC-PRP complex on TDSCs and tendinopathy. The aim of this study was to investigate the effect of BMAC-PRP on the TDSCs and to find the therapeutic effect of BMAC-PRP on the rotator cuff tendon tear. The chondrogenic and osteogenic potential of TDSCs decreased, but the adipogenic potential of TDSCs revealed no significant difference when they were cocultured with BMAC-PRP. Cell proliferation was significantly greater in TDSCs cocultured with BMAC-PRP than in TDSCs. The degree of wound closure (percentage) was different between TDSCs and TDSCs with BMAC-PRP. There was no significant difference in expression of collagen type I and type III in immunocytochemical staining in the presence of BMAC-PRP. Initial visual analog scale (VAS) score was 5.8 ± 1.9, which changed to 5.0 ± 2.3 at 3 weeks and 2.8 ± 2.3 at 3 months after the BMAC-PRP injection (p < 0.01). The American Shoulder Elbow Surgeon score changed from 39.4 ± 13.0 at baseline to 52.9 ± 22.9 at 3 weeks and 71.8 ± 19.7 at 3 months after the injection (p < 0.01). The initial torn area of the rotator cuff tendon was 30.2 ± 24.5 mm2, and this area was reduced to 22.5 ± 18.9 mm2 at 3 months, but the change was not significant (p > 0.05). The data indicate that BMAC-PRP enhances the proliferation and migration of TDSCs and prevents the aberrant chondrogenic and osteogenic differentiation of TDSCs, which might provide a mechanistic basis for the therapeutic benefits of BMAC-PRP for rotator cuff tendon tear.

Anti-inflammatory and lymphangiogenetic effects of low-level laser therapy on lymphedema in an experimental mouse tail model.

The aim of the present study was to investigate the therapeutic mechanism of low-level laser therapy (LLLT) in the mouse tail lymphedema model. Six-week-old female mice were classified into the laser treatment group, sham treatment group, and surgical control group (10 mice per group). LLLT was administered daily for 10 min from the surgical day to 11 days (12 times). Macrophage activation and lymphatic vessel regeneration were evaluated through immunohistochemical staining with anti-F4/80 and anti-LYVE-1 antibodies, respectively, at 12 days post-procedure. Quantitative real-time polymerase chain reaction (qPCR) was performed to measure messenger RNA (mRNA) expression of vascular endothelial growth factor A, B, C, R1, R2, and R3 (VEGF-A, VEGF-B, VEGF-C, VEGFR1, VEGFR2, and VEGFR3) at 12 days post-procedure. Student's t and one-way ANOVA tests were performed for statistical analyses. Significance was defined as p < 0.05. The thickness of the tail rapidly increased until 6 days in the laser and sham groups. The mice in the laser group showed a significantly decreased thickness compared with the sham group at 10 and 12 days. Immunohistochemistry assay revealed that LLLT reduced inflammation and induced new lymphatic vessel growth. qPCR showed that expressions of VEGFR3 were (p = 0.002) increased in the laser group. These results suggest that LLLT has anti-inflammatory and lymphangiogenetic effects for the management of lymphedema.

Atrazine induces endoplasmic reticulum stress-mediated apoptosis of T lymphocytes via the caspase-8-dependent pathway.

Atrazine (ATR) is one of the most commonly applied broad-spectrum herbicides. Although ATR is well known to be a biologically hazardous molecule with potential toxicity in the immune system, the molecular mechanisms responsible for ATR-induced immunotoxicity remain unclear. In this study, we found that the immunotoxic properties of ATR were mediated through the induction of apoptotic changes in T lymphocytes. Mice exposed to ATR for 4 weeks exhibited a significant decrease in the number of spleen CD3(+) T lymphocytes, while CD19(+) B lymphocytes and nonlymphoid cells were unaffected. ATR exposure also led to inhibition of cell growth and induction of apoptosis in human Jurkat T-cells. Importantly, ATR triggered the activation of caspase-3 and the cleavage of caspase-8 and PARP, whereas it did not affect the release of cytochrome c from the mitochondria in Jurkat T-cells. In addition, ATR activated the unfolded protein response signaling pathway, as indicated by eIF2α phosphorylation and CHOP induction. Our results demonstrate that ATR elicited an immunotoxic effect by inducing ER stress-induced apoptosis in T-cells, therefore providing evidence for the molecular mechanism by which ATR induces dysregulation of the immune system. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 998-1008, 2016.

High level of interleukin-32 gamma in the joint of ankylosing spondylitis is associated with osteoblast differentiation.

BACKGOUND: The formation of bony spurs and ankylosis is a key pathognomic feature in ankylosing spondylitis (AS) and results in functional impairment. The aim of this study was to investigate the role of IL-32γ in osteoblast (OB) differentiation and its association with the pathogenesis of AS. METHODS: The concentration and expression of IL-32γ were evaluated in synovial fluid and tissue from patients with AS, rheumatoid arthritis (RA) and osteoarthritis (OA), using enzyme-linked immunosorbent assay and immunohistochemistry. To establish whether IL-32γ affects OB differentiation, we used calvarial cells of IL-32γ transgenic (TG) mice or wild-type (WT) mice. To elucidate the mechanism of osteoblastogenesis, levels of regulators were assayed in IL-32γ TG mice and in primary OBs after IL-32γ stimulation. RESULTS: The IL-32γ levels were higher in the synovial fluid of AS patients compared with RA or OA patients and the expression of IL-32 was higher in AS synovia than in RA or OA synovia. Additional IL-32γ stimulation in precursor cells enhanced OB differentiation potentially and IL-32γ TG mice showed higher rates of OB differentiation than WT mice. IL-32γ reduced the expression of DKK-1, a negative regulator, in both WT precursor cells and human OBs and the constitutive expression of DKK-1 was suppressed in calvarial cells from IL-32γ TG mice. CONCLUSIONS: The elevated level of IL-32γ in AS joint could enhance OB differentiation via DKK-1 suppression. Therefore, IL-32γ might be a putative molecular target to prevent the abnormal bone formation in AS.

Interleukin-1ß promotes the LC3-mediated secretory function of osteoclast precursors by stimulating the Ca²âº-dependent activation of ERK.

Bone resorption by osteoclasts requires the release of secretory lysosomes containing cathepsin K, which degrades the organic bone matrix. The activity of this secretory function is determined by the level of lipidation of microtubule-associated protein 1 light chain 3 (LC3). Although the inflammatory cytokine IL-1ß increases osteoclast activity, the underlying mechanism(s) remains undefined. In our present study, we found that IL-1ß accelerates the release of cathepsin K from osteoclast precursors by increasing the cleavage and lipidation of LC3 and the subsequent formation of lipid-bound LC3-II containing secretory lysosomes. IL-1ß increased LC3-II formation within osteoclast precursors through a process that is dependent on increases in the intracellular Ca(2+) levels. In addition, IL-1ß was found to act synergistically with RANKL to increase ERK activation in a Ca(2+)-dependent manner. More importantly, Atg7-deficient osteoclast precursors, which showed impaired lipidation of LC3-I, did not exhibit IL-1ß-mediated increases in cathepsin K secretion. Thus, IL-1ß promotes LC3-II formation through the Ca(2+)-dependent activation of ERK, which triggers the release of cathepsin K. These findings provide evidence for a molecular mechanism through which IL-1ß enhances the secretory function of osteoclast precursors.

Secretory clusterin inhibits osteoclastogenesis by attenuating M-CSF-dependent osteoclast precursor cell proliferation.

Secretory clusterin (sCLU)/apolipoprotein J is a multifunctional glycoprotein that is ubiquitously expressed in various tissues. Reduced sCLU in the joints of patients with bone erosive disease is associated with disease activity; however, its exact role has yet to be elucidated. Here, we report that CLU is expressed and secreted during osteoclastogenesis in mouse bone marrow-derived macrophages (BMMs) that are treated with receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). CLU-deficient BMMs obtained from CLU(-/-) mice exhibited no significant alterations in OC differentiation in comparison with BMMs obtained from wild-type mice. In contrast, exogenous sCLU treatment significantly inhibited OC formation in both BMMs and OC precursor cultures. The inhibitory effect of sCLU was more prominent in BMMs than OC precursor cultures. Interestingly, treating BMMs with sCLU decreased the proliferative effects elicited by M-CSF and suppressed M-CSF-induced ERK activation of OC precursor cells without causing apoptotic cell death. This study provides the first evidence that sCLU reduces OC formation by inhibiting the actions of M-CSF, thereby suggesting its protective role in bone erosion.

Beclin-1 is required for RANKL-induced osteoclast differentiation.

Beclin-1 plays a critical role in autophagy; however, it also contributes to other biological processes in a non-autophagic manner. Although studies have examined the non-autophagic role of autophagy proteins in the secretory function of osteoclasts (OC), the role of Beclin-1 is unclear. Here, we examined the role of Beclin-1 in OC differentiation, and found that mouse bone marrow macrophages (BMMs) showed increased expression of Beclin-1 upon RANKL stimulation in a p38- and NF-kappa B-dependent manner. During OC differentiation, Beclin-1 localized to the mitochondria, where it was involved in the production of mitochondrial intracellular reactive oxygen species. Knockdown of Beclin-1 in RANKL-primed BMMs led to a significant reduction in RANKL-dependent osteoclastogenesis, which was accompanied by reduced NFATc1 induction. Furthermore, knockdown of Beclin-1 inhibited RANKL-mediated activation of JNK and p38, both of which act downstream of reactive oxygen species, resulting in the suppression of NFATc1 induction. Finally, overexpression of constitutively active NFATc1 rescued the phenotype induced by Beclin-1 knockdown, indicating that Beclin-1 mediates RANKL-induced osteoclastogenesis by regulating NFATc1 expression. These findings show that Beclin-1 plays a non-autophagic role in RANKL-induced osteoclastogenesis by inducing the production of reactive oxygen species and NFATc1.

PTX3 stimulates osteoclastogenesis by increasing osteoblast RANKL production.

Pentraxin-3 (PTX3), also known as tumor necrosis factor-stimulated gene 14 (TSG-14), is produced by immune and vascular cells in response to pro-inflammatory signals and is therefore a multipotent inflammatory mediator. The present study showed that during human osteoblast (OB) differentiation, precursor OBs (pOBs), but not mature OB, highly expressed PTX3. TNFα treatment elevated the PTX3 expression of pOBs. When mice were injected with lipopolysaccharide, which induces an inflammatory osteolytic condition characterized by trabecular bone destruction and high osteoclastogenesis, their bone marrow cells expressed elevated levels of PTX3 protein. Exogenous PTX3 did not directly affect osteoclast (OC) or OB differentiation. However, when pOBs and precursor OCs were co-cultured, exogenous PTX3 significantly increased the number of tartrate-resistant acid phosphatase-positive multinucleated cells (i.e., OC cells) by increasing the pOB mRNA expression and protein secretion of RANK ligand (RANKL). This was accompanied with increased Runt-related transcription factor 2 (Runx2) expression in the pOBs. Knock-down of endogenous PTX3 with small-interfering RNA did not change the osteogenic potential of pOBs but suppressed their production of RANKL and reduced osteoclastogenesis. Finally, TNFα treatment of the co-culture elevated PTX3 expression by the pOBs and increased OC formation. This effect was suppressed by PTX3 knock-down by decreasing RANKL expression. Thus, the PTX3-driven increase in the osteoclastogenic potential of pOBs appears to be mediated by the effect of PTX3 on pOB RANKL production. These findings suggest that PTX3 is an inflammatory mediator that contributes to the deteriorating osteolytic condition of inflamed bone. J. Cell. Physiol. 229: 1744-1752, 2014. © 2014 Wiley Periodicals, Inc.

Elevated Pentraxin 3 in bone metastatic breast cancer is correlated with osteolytic function.

Pentraxin 3 (PTX3), a modulator of tumor-associated inflammation, is known to be positively correlated with tumor grade and severity of malignancies, but its exact role remains unclear. This study found that PTX3 expression was up-regulated in distant bone metastases of breast cancer compared to lung, liver, and brain metastases in 64 human breast cancer patients. Elevated expression of PTX3 was correlated with poor survival in patients with breast cancer. PTX3 expression was also up-regulated in a bone metastatic breast cancer cell line and further enhanced by pro-inflammatory cytokine TNFα. Administration of PTX3 promoted the migratory potential of breast cancer cells and the mobilization of macrophages, a precursor of osteoclasts (OCs), toward breast cancer cells. In addition, elevated expression of PTX3 by TNFα led to enhanced OC formation, implying the distinct role of PTX3 in osteolytic bone metastasis of breast cancer cells. Furthermore, PTX3 silencing using PTX3-specific siRNA prevented breast cancer cell migration, macrophage chemotaxis, and subsequent OC formation. These findings provide an important insight into the key role of PTX3 in inflammation-associated osteolytic complications of breast cancer.

Multistep enzymatic synthesis of long-chain α,ω-dicarboxylic and ω-hydroxycarboxylic acids from renewable fatty acids and plant oils.

A multistep enzyme catalysis was successfully implemented to produce long-chain α,ω-dicarboxylic and ω-hydroxycarboxylic acids from renewable fatty acids and plant oils. Sebacic acid as well as ω-hydroxynonanoic acid and ω-hydroxytridec-11-enoic acid were produced from oleic and ricinoleic acid.