Osteoclasts (OCs), as the primary cells responsible for bone resorption, play a crucial role in the development of osteoporosis. Yet, how OC differentiation is regulated and involved in the occurrence of OP remains unknown. Here, we notice that BPAG1 is closely related to OC differentiation and osteoporosis. The upregulation of BPAG1 promotes OC differentiation in vitro, whereas its downregulation inhibits this process. Research on the mechanism indicates that BPAG1 can induce OC differentiation via regulating the phosphorylation of ERK. Notably, inhibiting ERK impaired the increase in OC differentiation caused by BPAG1 overexpression. In addition, inhibiting BPAG1 reversed bone destruction in osteoporotic mice. Hence, our research revealed the mechanism by which BPAG1 regulates OC differentiation and bone resorption by activating ERK. Targeting BPAG1 is a potential strategy for treating osteoporosis.

The vascular and lymphatic systems are integral to maintaining skeletal homeostasis and responding to pathological conditions in bone and joint tissues. This review explores the interplay between blood vessels and lymphatic vessels in bones and joints, focusing on their roles in homeostasis, regeneration, and disease progression. Type H blood vessels, characterized by high expression of CD31 and endomucin, are crucial for coupling angiogenesis with osteogenesis, thus supporting bone homeostasis and repair. These vessels facilitate nutrient delivery and waste removal, and their dysfunction can lead to conditions such as ischemia and arthritis. Recent discoveries have highlighted the presence and significance of lymphatic vessels within bone tissue, challenging the traditional view that bones are devoid of lymphatics. Lymphatic vessels contribute to interstitial fluid regulation, immune cell trafficking, and tissue repair through lymphangiocrine signaling. The pathological alterations in these networks are closely linked to inflammatory joint diseases, emphasizing the need for further research into their co-regulatory mechanisms. This comprehensive review summarizes the current understanding of the structural and functional aspects of vascular and lymphatic networks in bone and joint tissues, their roles in homeostasis, and the implications of their dysfunction in disease. By elucidating the dynamic interactions between these systems, we aim to enhance the understanding of their contributions to skeletal health and disease, potentially informing the development of targeted therapeutic strategies.

Schwann cells (SCs) are the major component of myelin sheath in the peripheral nervous system, which are necessary in the development, function maintenance, and repair of peripheral nerves. This study aimed to investigate the potential mechanism of low-intensity pulsed ultrasound (LIPUS) affecting the proliferation and myelinating activity of SCs. Rat Schwann cell line RSC96 were cultured and exposed to LIPUS of different duty ratios (control, 20 %, 50 %, 80 %). Results demonstrated that LIPUS with a duty ratio of 50 % showing the maximal effect in facilitating proliferation of SCs. The expressions of Krox20 and myelin basic protein (MBP), the key molecules of SC myelination, and the potent inducer of myelination neuregulin 1 (NRG1) and its receptors ErbB2 and ErbB3 increased significantly by LIPUS. The reaction of these factors to LIPUS were both time- and duty ratio-dependent: namely LIPUS with higher duty ratios took effects when applied repeatedly over more consecutive days. These observations indicated that NRG1/ErbB signaling pathway might contribute to the effects of LIPUS on the proliferation and myelinating status of SCs, which could be one of the mechanisms in the protective role of LIPUS in nerve repair and regeneration. Our work provided novel insights for promising strategies of nerve repair therapy.

Response surface technique was employed for improving the extraction of corn silk polysaccharides (CSP). Temperature, liquid-to-solid ratio, and per extraction time were all examined as separate factors. The optimal extraction parameters were determined by fitting experimental data to a second-order polynomial; a liquid-to-solid ratio of 21.5 ml/g, temperature equivalent to 88°C, and extraction time of 1.87 h. The experimental yield of the extracted polysaccharides following the application of these conditions was 4.33 ± 0.08% (dry weight), which fit quite well with the predicted value. CSP’s strong scavenging capabilities against hydroxyls, 1,1-diphenyl-2-picrylhydrazyl radicals, and superoxide anions along with its excellent reducing potential, were demonstrated in an in vitro antioxidant experiment. Meanwhile, in vivo testing revealed that CSP substantially enhanced glutathione peroxidase and superoxide dismutase activities. The Malondialdehyde levels in the liver and serum of aged mice also underwent a decrease. This study found that CSP has a substantial antioxidant potential in vitro and in vivo, suggesting that it might be used as an antioxidant in food and medicine.

OBJECTIVE:This study aimed to explore the effect of Low-intensity pulsed ultrasound (LIPUS) on implant osseointegration and elucidate the role of α-calcitonin gene-related peptide (αCGRP) in this process. DESIGN:In vivo, αCGRP+/+ (Wild-type model) mice and αCGRP-/- (Knock-out model) mice with implants immediately placed in the maxillary first molars extraction sockets were treated with LIPUS. We detected details of peri-implant bone tissues by micro-CT, real-time PCR and histological analysis. In vitro, αCGRP+/+ and αCGRP-/- dorsal root ganglia (DRG) neurons were cultured and exposed to LIPUS. Then conditioned media from these neurons were collected and added to osteoblasts to analyze cell differentiation, mineralization and proliferation by real-time PCR, alkaline phosphatase (ALP) and cell counting kit-8 (CCK-8) assay. Besides, ELISA was performed to determine the effect of LIPUS on the αCGRP secretion in neurons. RESULTS:In vivo tests revealed that αCGRP-/- mice displayed worse osseointegration when compared to αCGRP+/+ mice. LIPUS could enhance implant osseointegration in αCGRP+/+ mice but had little effect on αCGRP-/- mice. Meanwhile, αCGRP was elevated during the osseointegration with LIPUS treatment. In vitro, LIPUS promoted αCGRP secretion in DRG neurons, thereby enhanced osteogenic differentiation and mineralization of osteoblasts. Also we proved that the effects of LIPUS was duty cycle-related and LIPUS of 80% duty cycle had the strongest impacts. CONCLUSIONS:Our findings demonstrated that LIPUS could enhance osseointegration of dental implant by inducing local neuronal production of αCGRP, providing a new idea to promote peri-implant osseointegration and bone regeneration.

Low-intensity pulsed ultrasound (LIPUS) is a common physical therapy to accelerate the healing of bone fracture and treat delayed union of bone fracture. Vessels, nerves, and bone tissue are essential constituents of bone system. Recently, increasing evidence has been revealed that LIPUS can not only promote bone regeneration by directly regulating osteoblasts, osteoblasts, mesenchymal stem cells, but also have a positive impact on the repair of bone healing through vessels and nerves. Thus, we reviewed and summarized the latest published literature about the molecular mechanism for the effects of LIPUS on bone regeneration, which might offer a promising therapy for bone-related diseases.

This study is aimed at investigating the effect of amifostine (AMI) on rat bone marrow stromal stem cells (BMSCs) exposed to 2 Gy radiation. The BMSCs were divided into four groups, namely, group A that received 0 Gy radiation, group B that received 0 Gy radiation and AMI, group C that received 2 Gy radiation, and group D that received 2 Gy radiation and AMI. The proliferation, apoptosis, and distribution of BMSCs in the cell cycle, along with their osteogenesis ability, adipogenesis ability, and ROS production, were subsequently examined. The levels of ALP, PPAR γ , P53, and TNF α were determined by Western blotting. The results demonstrated that the proliferation of BMSCs and the levels of ALP in group C were much lower than those in group A. The production of ROS and levels of PPAR γ , P53, and TNF α in the group that received 2 Gy radiation were much higher than those in group A. Furthermore, the production of ROS and the levels of PPAR γ , P53, and TNF α were much lower in group D than in group C. Additionally, the levels of ALP and extent of cell proliferation were much higher in group D than in group C. The results demonstrated the potential of AMI in reducing the side effects of radiation in BMSCs and in treatment of bone diseases caused by radiation.

Patients with malignant tumors receive radiotherapy, and radiation could harm the skeletal system, leading to radiation-induced osteoporosis. A major cause of this phenomenon is the activation of osteoclasts by radiotherapy. In this study, we studied whether amifostine (AMI) could affect the differentiation of osteoclast precursor cells (RAW264.7 cells) into osteoclasts under 2 gray (Gy) radiation. Four groups were used in the experiment: (a) 0 Gy (no radiation); (b) 0 Gy + AMI; (c) 2 Gy radiation; and (d) 2 Gy radiation + AMI. After radiation, a proliferation assay, a reactive oxygen species (ROS) assay, a comet assay, Trap staining, reverse transcription polymerase chain reaction, and an animal study to test the effect of AMI on osteoclast precursor cells under 2 Gy radiation were conducted. Cell proliferation was significantly inhibited by AMI (P < .05). In addition, 2 Gy radiation led to longer "comet tails", high level of ROS, and more Trap-positive cells in vivo and in vitro (P < .05). Radiation improved the expression of CSTK, NFAT, and Rankl/OPG gene (P < .05), as well as Trap-5b levels in the serum, and decreased bone mineral density. AMI inhibited the differentiation of RAW264.7 cells, shortened the tail moment length of comets, and decreased the level of ROS induced by radiation. The expression of NFAT, CTSK, and Rankl/OPG was decreased by AMI at the detection time point in radiation groups (P < .05). AMI inhibits the maturation and differentiation of osteoclasts under radiation conditions by reducing DNA damage and ROS induced by radiation, thereby reducing the adverse effects of radiation in the skeletal system, indicating that AMI might be used to treat osteoradionecrosis.

The present study aimed to elucidate whether extracellular signal-regulated kinases 1/2 (ERK1/2) and p38 mitogen-activated protein kinases pathways participate in the transduction of mechanical stretch exerted on adipose stem cells (ASCs) into intracellular osteogenic signals, and if so whether both pathways have time-dependent feature. Rat ASCs were cultured in osteogenic medium for 72 h and assigned into three sets, namely ERK1/2 inhibitor treated set, p38 inhibitor treated set, and the control set. After inhibitor treatment, all cells were subjected to cyclic stretch(2000 με, 1 Hz) on a four-point bending mechanical loading device. Protein and mRNA samples were acquired at six time points: 0, 15 min, 30 min, 1 h, 2 h and 6 h. Western blot showed phosphorylation level of ERK1/2 was elevated by cyclic tensile stress at all time points, while p38 at 15 min, 30 min and 1 h, and the elevation can be completely blocked by corresponding inhibitors. The treatment by ERK1/2 inhibitor was shown to antagonize the up-regulation of osteogenic genes bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (Runx2) by mechanical stretch at 15 min and 6 h, whereas p38 inhibitor took effect at 15 min only. The results suggested both ERK and p38 could be positive mediators of stretch-induced osteogenic differentiation of ASCs, and ERK stimulate the stretch-induced osteogenic differentiation at both early and late stages while p38 responds to mechanical stretch in a more rapid fashion.

Objective To investigate the effects of low-intensity pulsed ultrasound (LIPUS) on the mandibular remodeling following inferior alveolar nerve transection (IANX) and to optimize the parameters of LIPUS in the treatment of nerve injury. Methods IANX was performed in male Sprague-Dawley rats. Four weeks after IANX,the effect of daily LIPUS (from day 1) on the transected inferior alveolar nerve was examined in terms of sensitivity to mechanical stimulation. Moreover,histopathologic changes of mandibles were analyzed by micro-CT,HE staining,Masson trichrome staining,and immunohistochemical staining. Results LIPUS promoted the recovery of inferior alveolar nerve injury after transection. HE staining displayed the improvement of trabecular thickness and continuity. LIPUS with higher duty ratios had more obvious effect on bone remodeling. Conclusion LIPUS promotes the mandibular remodeling following IANX.

OBJECTIVES:Peripheral nerve injuries are a common occurrence, resulting in considerable patient suffering; it also represents a major economic burden on society. To improve treatment options following peripheral nerve injuries, scientists aim to find a way to promote Schwann cell (SC) myelination to help nerves to carry out their functions effectively. In this study, we investigated myelination ability of SCs, regulated by co-culture with adipose-derived stem cells (ASCs) or low-intensity pulsed ultrasound (LIPUS), and synergistic effects of combined treatments. MATERIALS AND METHODS:Schwann cells were co-cultured with or without ASCs, and either left untreated or treated with LIPUS for 10 min/d for 1, 4 or 7 days. Effects of LIPUS and ASC co-culture on pro-myelination indicators of SCs were analysed by real-time PCR (RT-PCR), Western blotting and immunofluorescence staining (IF). RESULTS:Our results indicate that ASC-SC co-culture and LIPUS, together or individually, promoted mRNA levels of epidermal growth factor receptor 3 (EGFR3/ErbB3), neuregulin1 (NRG1), early growth response protein 2 (Egr2/Krox20) and myelin basic protein (MBP), with corresponding increases in protein levels of ErbB3, NRG1 and Krox20. Interestingly, combination of ASC-SC co-culture and LIPUS displayed the most remarkable effects. CONCLUSION:We demonstrated that ASCs upregulated pro-myelination indicators of SCs by indirect contact (through co-culture) and that effects could be potentiated by LIPUS. We conclude that LIPUS, as a mechanical stress, may have potential in nerve regeneration with potential clinical relevance.

OsMADS32 is a monocot specific MIKC(c) type MADS-box gene that plays an important role in regulating rice floral meristem and organs identity, a crucial process for reproductive success and rice yield. However, its underlying mechanism of action remains to be clarified. Here, we characterized a hypomorphic mutant allele of OsMADS32/CFO1, cfo1-3 and identified its function in controlling rice flower development by bioinformatics and protein-protein interaction analysis. The cfo1-3 mutant produces defective flowers, including loss of lodicule identity, formation of ectopic lodicule or hull-like organs and decreased stamen number, mimicking phenotypes related to the mutation of B class genes. Molecular characterization indicated that mis-splicing of OsMADS32 transcripts in the cfo1-3 mutant resulted in an extra eight amino acids in the K-domain of OsMADS32 protein. By yeast two hybrid and bimolecular fluorescence complementation assays, we revealed that the insertion of eight amino acids or deletion of the internal region in the K1 subdomain of OsMADS32 affects the interaction between OsMADS32 with PISTILLATA (PI)-like proteins OsMADS2 and OsMADS4. This work provides new insight into the mechanism by which OsMADS32 regulates rice lodicule and stamen identity, by interaction with two PI-like proteins via its K domain.

BACKGROUND:Successful drug treatment for ischemia--reperfusion-induced lung injury remains a major clinical problem. Melatonin (MT) is a hormone that is principally synthesized in the pineal gland. It has been shown to exhibit a variety of functions including anti-inflammatory and antioxidant effects. Previous reports on N-myc downstream-regulated gene (NDRG)2 have suggested that it is involved in cellular differentiation, development, antiapoptosis, anti-inflammatory cytokine, and antioxidant. The objective of this study was to test whether MT, a novel NDRG2 activator, can protect against intestinal ischemia-reperfusion-induced lung injury (IIRI). MATERIALS AND METHODS:IIRI was induced in rats by occlusion of the superior mesenteric artery for 60 min, and the occlusion was then released for reperfusion. Rats were randomly divided into six groups as follows: control group; MT group; IIRI group; IIRI+5 mg/kg MT group; IIRI+15 mg/kg MT group; and IIRI+25 mg/kg MT group. The effects of MT on intestinal ischemia-reperfusion-induced lung pathologic changes, inflammatory cytokines release, myeloperoxidase and superoxide dismutase activities, and malondialdehyde level were examined. In addition, the NDRG2 activation in lung tissues was detected by Western blot analysis. RESULTS:MT pretreatment attenuated edema and the pathologic changes in the lung. MT also decreased the levels of tumor necrosis factor-α, interleukin-1β, and interleukin-8 in bronchoalveolar lavage fluid. In addition, MT markedly prevented IIRI-induced elevation of malondialdehyde and myeloperoxidase levels, as well as reduction of superoxide dismutase activity. Furthermore, the expression of NDRG2 was activated by MT pretreatment in lung tissues. CONCLUSIONS:The present study demonstrates that MT exerted protection against IIRI-induced oxidative stress. The potential mechanism of this action may attribute partly to the activation of NDRG2 expression.

Mechanical forces play critical roles in the development and remodeling processes of bone.As an alternative cell source for bone engineering,adipose-derived stem cells（ASCs）should be fully investigated for their responses to mechanical stress.Similarly,the osteogenic potential,stimulated by mechanical stress,should be compared with bone marrow stromal cells（BMSCs）,which have been clinically used for bone tissue engineering.In this study,ASCs and BMSCs were osteogenic-induced for 48 hours,and then subjected to uniaxial mechanical stretching for 2 or 6 hours.Cell orientation,osteogenic regulatory genes,osteogenic genes and ALP activities were measured and compared between ASCs and BMSCs.ASCs could align in a perpendicular way to the direction of stretching stress,while BMSCs did not present a specific alignment.Both 2 and 6 hours mechanical stretching could enhance the mRNA expression of Osx and Runx2 in BMSCs and ASCs,while OCN mRNA only increased in ASCs after 6 hours mechanical loading.Mechanical stretching enhanced the BMP-2 mRNA expression in ASCs,while only after 6 hours of mechanical loading significantly increased the BMP-2 gene expression in BMSCs.Significant differences only exist between ASCs and BMSCs loaded at 2 hours of mechanical stretching.It is concluded that ASCs are more rapid responders to mechanical stress,and have greater potential than BMSCs in osteogenesis when stimulated by mechanical stretching,indicating their usefulness for bone study in a rat model.

Based on the highly conserved sequences of other plants,counterpart sequences of CHS (chalcone synthase,CHS)and LDOX(leucoanthocyanidin dioxygenase,LDOX)of Actinida chinesis,'Hongyang'were obtained using the special primers.Full-length cDNAs encoding CHS and LDOX were cloned respectively from fruit of'Hongyang'by RACE(rapid amplification of cDNA ends).The AcCHS was 1 501 bp in length,encoding 389 amino acids.AcLDOX was 1 381 bp in length and encoded 355 amino acids.Amino acids sequence of AcCHS exhibited over 95% homology with CHS of Abelmoschus manihot,Camellia japonica and Gossypium hirsutum.It also shared 94% and 93% homology with Vitis vinifera and Malus × domestica respectively.AcLDOX showed 94% and 93% identity with Vitis amurensis and Vitis vinifera respectively at amino acids level.The expression of AcCHS and AcLDOX in the inner pericarp of three kiwifruit cultivars with red,green and yellow color was analyzed by Real-time PCR.AcCHS expressed highly in flesh of'Hongyang'at 65 DAF(days after flower).Expression of the gene in yellow-fleshed'Jinnong'decreased permanently after flowering.Expression of AcLDOX increased during the early developmental stage in'Hongyang',but dropped quickly after 65 DAF.Interestingly,expression of AcLDOX raised markedly in the late developmental stages of green-fleshed'Jinkui',which was the highest among three cultivars.AcLDOX in'Jinnong'also deregulated after flowering.

The clinical use of dental implants has a high success rate, but overload-induced bone loss around implant is not uncommon in patients with implant-supported denture especially those with long cantilever designs and greatly harmful to the long-term implant success. The mechanism underlying the bone loss is thought to be the imbalance of bone remodeling involving a detrimental positive feedback activated by overloading. While surgical regenerative treatments may be useful in promoting bone regeneration, extra suffering and risk of infection have to be fully recognized. To date, no optimal method is available to solve this problem. We hereby propose a novel therapy that may potentially improve this condition. Many studies have shown that parathyroid hormone (PTH), an anabolic agent targeting bone, is effective in reversing bone loss caused by osteoporosis with negative bone remodeling in clinical studies. Moreover, PTH has the potential to accelerate the bone healing in patients with fracture and fracture nonunion and improve osseointegration of implant inserted in pre-existed bone defect via its anabolic effect to increase bone formation in animal model studies. Specifically for alveolar bone, PTH is associated with effective bone regeneration in patients with severe periodontitis. What is more, PTH and mechanical loading has a synergistic effect on bone formation, which is in favor of bone healing under physiological loading. The mechanisms underlying its anabolic effect may involve increased osteoblasts activity, prolonged osteoblasts life-span and recruitment of new osteoblasts from marrow stromal cells. Furthermore, PTH could activate resting lining cells to initial de novo bone formation. Considering these actions of PTH, we hypothesize that PTH may be a potential treatment for overload-induced bone loss around implant.

The original article to which this Retraction refers was published in J Cell Biochem 2006; DOI 10.1002/jcb.21155. © 2006 Wiley‐Liss, Inc..