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1.
FASEB J ; 38(13): e23776, 2024 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-38958998

RESUMO

This study aimed to explore how mechanical stress affects osteogenic differentiation via the miR-187-3p/CNR2 pathway. To conduct this study, 24 female C57BL/6 mice, aged 8 weeks, were used and divided into four groups. The Sham and OVX groups did not undergo treadmill exercise, while the Sham + EX and OVX + EX groups received a 8-week treadmill exercise. Post-training, bone marrow and fresh femur samples were collected for further analysis. Molecular biology analysis, histomorphology analysis, and micro-CT analysis were conducted on these samples. Moreover, primary osteoblasts were cultured under osteogenic conditions and divided into GM group and CTS group. The cells in the CTS group underwent a sinusoidal stretching regimen for either 3 or 7 days. The expression of early osteoblast markers (Runx2, OPN, and ALP) was measured to assess differentiation. The study findings revealed that mechanical stress has a regulatory impact on osteoblast differentiation. The expression of miR-187-3p was observed to decrease, facilitating osteogenic differentiation, while the expression of CNR2 increased significantly. These observations suggest that mechanical stress, miR-187-3p, and CNR2 play crucial roles in regulating osteogenic differentiation. Both in vivo and in vitro experiments have confirmed that mechanical stress downregulates miR-187-3p and upregulates CNR2, which leads to the restoration of distal femoral bone mass and enhancement of osteoblast differentiation. Therefore, mechanical stress promotes osteoblasts, resulting in improved osteoporosis through the miR-187-3p/CNR2 signaling pathway. These findings have broad prospect and provide molecular biology guidance for the basic research and clinical application of exercise in the prevention and treatment of PMOP.


Assuntos
Diferenciação Celular , MicroRNAs , Osteoblastos , Osteogênese , Osteoporose Pós-Menopausa , Estresse Mecânico , Animais , Feminino , Humanos , Camundongos , Células Cultivadas , Camundongos Endogâmicos C57BL , MicroRNAs/genética , MicroRNAs/metabolismo , Osteoblastos/metabolismo , Osteoporose Pós-Menopausa/metabolismo , Osteoporose Pós-Menopausa/terapia , Osteoporose Pós-Menopausa/genética , Osteoporose Pós-Menopausa/patologia , Transdução de Sinais
2.
Small ; 20(26): e2309868, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38259052

RESUMO

Critical-sized segmental long bone defects represent a challenging clinical dilemma in the management of battlefield and trauma-related injuries. The residual bone marrow cavity of damaged long bones contains many bone marrow mesenchymal stem cells (BMSCs), which provide a substantial source of cells for bone repair. Thus, a three-dimensional (3D) vertically aligned nanofiber scaffold (VAS) is developed with long channels and large pore size. The pore of VAS toward the bone marrow cavity after transplantation, enables the scaffolds to recruit BMSCs from the bone marrow cavity to the defect area. In vivo, it is found that VAS can significantly shorten gap distance and promote new bone formation compared to the control and collagen groups after 4 and 8 weeks of implantation. The single-cell sequencing results discovered that the 3D nanotopography of VAS can promote BMSCs differentiation to chondrocytes and osteoblasts, and up-regulate related gene expression, resulting in enhancing the activities of bone regeneration, endochondral ossification, bone trabecula formation, bone mineralization, maturation, and remodeling. The Alcian blue and bone morphogenetic protein 2 (BMP-2) immunohistochemical staining verified significant cartilage formation and bone formation in the VAS group, corresponding to the single-cell sequencing results. The study can inspire the design of next-generation scaffolds for effective long-bone regeneration is expected by the authors.


Assuntos
Regeneração Óssea , Diferenciação Celular , Condrogênese , Células-Tronco Mesenquimais , Nanofibras , Osteogênese , Alicerces Teciduais , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Nanofibras/química , Alicerces Teciduais/química , Animais
3.
Chin J Traumatol ; 2024 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-39242245

RESUMO

PURPOSE: Autologous osteoperiosteal transplantation (AOPT) is one of the most feasible and effective techniques for cystic osteochondral lesions of the talus (OLT). However, few reports have been reported about the process of graft-host bone healing and bone articular surface reconstruction, which help us to further understand the actual situation of bone healing and modify surgical methods. METHODS: We retrospectively evaluated 33 osteochondral lesions in 30 patients undertaking AOPT for OLT with subchondral cysts from December 2016 to October 2021. According to CT observation, we used 4 variables to describe the bony articular repair, including the integration of the articular surface, the height of the bone filling, the status of bone union, and the appearance of bone resorption or cystic change. We also analyzed the demographic data and clinical function. Descriptive statistics were used for demographic and clinical variables. Normally distributed data were presented as mean ± SD, and non-normally distributed data were presented as median (Q1, Q3). Associations between these variables and the primary clinical outcomes were examined using t-test or one-way ANOVA test for continuous variables. RESULTS: The patients' mean age was (41.7 ± 14.0) years old and the mean follow-up time was (29.6 ± 17.8) months. The chondral lesion size was (14.3 ± 4.1) mm. The cyst depth was (10.9 ± 3.7) mm. Significant improvements were observed in functional outcomes (according to the numeric rating scale for pain when walking and the American orthopedic foot and ankle society score) between the preoperative and latest follow-up evaluations, from 4.2 ± 2.1 to 2.2 ± 2.0 (p < 0.001), and from 66.8 ± 12.9 to 83.2 ± 10.4, respectively (p < 0.001). The overall satisfaction reached 8.3 of 10 points. All patients returned to sports and their median daily steps reached 8000 steps with 27 (81.8%) patients walking over 6000 steps daily. According to CT observation, "discontinuous bony articular surface and gap > 1 mm" was found in 27 grafts (81.8%), and "below the level of the adjacent articular surface, ≤ 1 mm" in a third of the grafts. Abnormal height of bone filling affected numeric rating scale score (p = 0.049) and American Orthopedic Foot and Ankle Society score (p = 0.027). Of note, bone resorption or cystic changes appeared in up to 13 autografts (39.4%). CONCLUSIONS: AOPT is an effective and acceptable technique for cystic OLT. Bone reconstruction is essential for large cystic OLT. How to get better bony articular reconstruction and avoid cyst recurrence should still be paid more attention.

4.
Foot Ankle Surg ; 30(3): 239-244, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38123374

RESUMO

BACKGROUND: The authors developed a simple extensible external fixator, which has the advantages of easy application and inexpensiveness. The present study aimed to make a comparison between this external fixator and calcaneal traction in preoperative temporary fixation for malaligned ankle fractures and pilon fractures. METHODS: From May 2020 to February 2022, patients with malaligned ankle fractures or Rüedi-Allgöwer type 2 or 3 pilon fractures with obvious soft tissue swelling were retrospectively reviewed and divided into the calcaneal traction group and the external fixation group. The two groups of patients were matched 1:1 before making comparisons. RESULTS: A total of 38 patients were included. Higher General Comfort Questionnaire score and lower visual analog scale score were noticed in the external fixation group during hospitalization (p < 0.05), while the operation latency time, total cost, patient satisfaction, and functional outcomes one year after surgery were not significantly different between the two groups. No wound complications were observed. CONCLUSION: Preoperative temporary fixation for fractures around the ankle using this simple extensible external fixator significantly improves patient comfort when compared to calcaneal traction. LEVEL OF EVIDENCE: III, retrospective comparative study.


Assuntos
Fraturas do Tornozelo , Traumatismos do Tornozelo , Fraturas da Tíbia , Humanos , Estudos Retrospectivos , Fraturas do Tornozelo/cirurgia , Tornozelo , Conforto do Paciente , Fixação Interna de Fraturas , Resultado do Tratamento , Traumatismos do Tornozelo/cirurgia , Fraturas da Tíbia/cirurgia , Fixadores Externos
5.
Knee Surg Sports Traumatol Arthrosc ; 31(6): 2174-2182, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36515734

RESUMO

PURPOSE: To compare the clinical outcomes of the lasso-loop and simple suture techniques in arthroscopic anterior talofibular ligament (ATFL) repair for the treatment of chronic lateral ankle instability (CLAI). METHODS: From 2018 to 2020, patients with CLAI who underwent arthroscopic ATFL repair using the lasso-loop or simple suture technique were matched 1:1 (arthroscopic lasso-loop [AL] group, n = 29; simple arthroscopic suture [AS] group, n = 29) based on age, sex, affected side, body mass index, and follow-up duration using propensity score matching and retrospectively evaluated. Karlsson score, visual analogue scale (VAS) score, Tegner score, anterior drawer test (ADT) results, complications, patient-reported satisfaction, and magnetic resonance (MR) re-evaluation findings of ATFL quality were used to describe the outcomes. RESULTS: The patient characteristics or follow-up durations did not significantly differ between the two groups. The Karlsson score, VAS score, and Tegner score improved significantly in both groups after a mean follow-up duration of 29.6 ± 2.8 months. The postoperative clinical scores, ADT results, satisfaction rates, complication rates and MR re-evaluation findings were not significantly different between the two groups at the latest follow-up. CONCLUSION: The lasso-loop technique was equivalent to the simple suture technique in arthroscopic ATFL repair for the treatment of CLAI after a minimum follow-up of 2 years, suggesting that the simple suture technique is sufficient for arthroscopic ATFL repair in most patients without the need to add a lasso loop. LEVEL OF EVIDENCE: Level III.


Assuntos
Instabilidade Articular , Ligamentos Laterais do Tornozelo , Humanos , Articulação do Tornozelo/cirurgia , Estudos Retrospectivos , Artroscopia/métodos , Ligamentos Laterais do Tornozelo/cirurgia , Instabilidade Articular/cirurgia , Técnicas de Sutura
6.
Connect Tissue Res ; 63(4): 370-381, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-34355626

RESUMO

PURPOSE: This study aimed to elucidate the effect of bone marrow mesenchymal stem cell (BMSC) transplantation combined with the administration of Lugua polypeptide injection into the knee joint cavity to treat knee osteoarthritis (KOA) in rabbits. MATERIAL AND METHODS: Sixty white New Zealand rabbits were randomly divided into the blank, model, Lugua polypeptide, BMSC, and combined (Lugua polypeptide plus BMSC) groups, with 12 rabbits in each group. The mRNA and protein expression levels of cyclin D1, bcl-2, TIMP-1, p21, caspase-3, Bax, MMP-1, MMP-13, TLR-4, and NF-κB p65 in chondrocytes, and levels of IL-1, NO, TNF-α, and IL-6 in the synovial fluid were compared. RESULTS: The severity of cartilage damage in the combined group was significantly less (P <0.01). Compared to the MG, the mRNA and protein expression levels of cyclin D1, bcl-2 and TIMP-1 in chondrocytes of the three other groups were significantly increased, while those of p21, caspase-3, Bax, MMP-1, MMP-13, TLR-4, and NF-κB p65 in the chondrocytes and levels of IL-1, NO, TNF-α, and IL-6 in the synovial fluid of the three other groups were significantly reduced (P <0.05). The aforementioned indicators in the combined group were significantly better than those of the Lugua polypeptide and BMSCs groups (P <0.05). CONCLUSIONS: BMSC transplantation combined with Lugua polypeptide injection may improve KOA-related cartilage tissue damage in rabbits.


Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Osteoartrite do Joelho , Animais , Medula Óssea/metabolismo , Caspase 3 , Condrócitos/metabolismo , Ciclina D1 , Interleucina-1/farmacologia , Interleucina-6 , Articulação do Joelho/metabolismo , Metaloproteinase 1 da Matriz/farmacologia , Metaloproteinase 13 da Matriz/metabolismo , Células-Tronco Mesenquimais/metabolismo , NF-kappa B/metabolismo , Osteoartrite do Joelho/terapia , RNA Mensageiro , Coelhos , Inibidor Tecidual de Metaloproteinase-1/genética , Receptor 4 Toll-Like , Fator de Necrose Tumoral alfa/farmacologia , Proteína X Associada a bcl-2/farmacologia
7.
Bioorg Med Chem ; 76: 117098, 2022 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-36455508

RESUMO

Through structural optimization and ring fusion strategy, we designed a series of novel imidazo[1,2-a]pyrazine derivatives as potential tubulin inhibitors. These compounds displayed potent anti-proliferative activities (micromolar to nanomolar) against a panel of cancer cell lines (including HepG-2, HCT-116, A549 and MDA-MB-231 cells). Among them, compound TB-25 exhibited the strongest inhibitory effects against HCT-116 cells with an IC50 of 23 nM. Mechanism studies revealed that TB-25 could effectively inhibit tubulin polymerization in vitro, and destroy the dynamic equilibrium of microtubules in HCT-116 cells. In addition, TB-25 dose-dependently induced G2/M phase cell cycle arrest and apoptosis in HCT-116 cells. Furthermore, TB-25 suppressed HCT-116 cell migration in a concentration-dependent manner. Finally, molecular docking showed that TB-25 fitted well in the colchicine binding site of tubulin and overlapped nicely with CA-4. Collectively, these results suggest that TB-25 represents a promising tubulin inhibitor deserving further investigation.


Assuntos
Moduladores de Tubulina , Tubulina (Proteína) , Moduladores de Tubulina/farmacologia , Pirazinas/farmacologia , Simulação de Acoplamento Molecular
8.
Int J Med Sci ; 19(8): 1275-1289, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35928719

RESUMO

Changes in DRG after nerve injury involve neuronal damage, apoptosis, pain transmission, and activation of regenerative programs. It is unclear which genes and microRNAs may play a major role in this process. Therefore, this study performed a meta-analysis of previously published gene expression data to reveal the potential microRNA-mRNA network in dorsal root ganglia (DRG) after peripheral nerve injury. We searched 5 mRNA and 3 microRNA expression data sets, obtained 447 differentially expressed genes (DEGs) and 5 differentially expressed miRNAs, determined the biological pathways enriched by these DEGs, and further predicted new microRNA-mRNA interactions, such as miR-21/Hmg20a, miR-221/Ube2ql1, miR-30c-1/Rhoq, miR-500/Sema3c, and miR-551b/Cdc42se2. We verified these hub mRNA and miRNA in rats by qRT-PCR and found the results were consistent with the bioinformatics analysis. And we predicted transcription factors associated with these genes (gTFs) and TFs associated with these microRNAs (mTFs) and constructed the mTF-miRNA-gene-gTF regulatory network to further explore the molecular mechanism in DRG. Finally, we compared the DRG transcriptome after PNI to that of chronic constriction injury (CCI), and found that PNI caused greater damage to DRG compared to CCI. At the same time, the related mechanisms of pain caused by the two pathophysiological process may be different.


Assuntos
MicroRNAs , Traumatismos dos Nervos Periféricos , Animais , Gânglios Espinais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Dor , Traumatismos dos Nervos Periféricos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos
9.
J Foot Ankle Surg ; 61(2): 434-437, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34896009

RESUMO

It is technical demanding work to perform arthroscopic ankle arthrodesis to treat end-stage ankle osteoarthritis with excessive talar tilt. This article aimed to provide an effective technique tip for the treatment of Takakura stage 3b ankle osteoarthritis with a talar tilt angle more than 15 degrees under arthroscopy. A conventional anterior arthroscopic approach is used. After arthroscopic examination and debridement, one pin which is parallel to the distal tibial surface is inserted into the tibial side of the ankle, the other pin which is parallel to the talar dome surface is inserted into the talar side of the ankle, both at the coronal plane. Then a distracter is used in the medial side to open the interspace of the tibiotalar joint and correct the talar tilt through the 2 pins, under which circumstance the tibiotalar joint surface can be well prepared. Next an anti-distracter is used in the lateral side to close the tibiotalar interspace and correct the talar tilt through the 2 pins, in which condition 3 fully threaded cannulated lag screws can be inserted through guide pins in a cross pattern to fix the ankle joint. We used the pin-based distracter to open and close tibiotalar interspace, correct the talar tilt and maintain a good mechanical axis for fusion, and the outcomes were good.


Assuntos
Osteoartrite , Tálus , Tornozelo , Articulação do Tornozelo/diagnóstico por imagem , Articulação do Tornozelo/cirurgia , Artrodese/métodos , Humanos , Osteoartrite/diagnóstico por imagem , Osteoartrite/cirurgia , Tálus/diagnóstico por imagem , Tálus/cirurgia
10.
J Foot Ankle Surg ; 61(5): 1056-1059, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35193803

RESUMO

The traditional lateral "L" approach is common for managing calcaneal fractures with a drawback of significant blood loss. Yet there are no prospective studies on the hemostatic effect of the topical use of tranexamic acid (TXA) in calcaneal fracture surgeries. The purpose of this study was to evaluate the role of topical administration of TXA in reducing postoperative blood loss in calcaneal fractures. Forty participants were randomly distributed into the TXA group (n = 20) and the control group (n = 20). All participants underwent the same surgery via the lateral "L" approach. At the end of the operation, the surgical wound was irrigated with 80 mL 0.5 g/L TXA in the TXA group and 80 mL 0.9% sodium chloride in the control group, followed by the routine use of a drainage tube when closing the incision. Then, 20 mL 0.5 g/L TXA (TXA group) or 20 mL 0.9% sodium chloride solution (control group) was injected retrogradely into the wound through the drainage tube, which was clipped for 30 minutes thereafter. There were no significant differences in the baseline data between the 2 groups (p > .05). There was significantly less blood loss in the first 24 hours and total blood loss postoperation in the TXA group (p < .01). The surgical wounds healed well after surgery in both groups with no complication. We concluded that topical application of TXA in calcaneal fracture surgeries is a safe and useful method that can reduce postoperative blood loss.


Assuntos
Traumatismos do Tornozelo , Antifibrinolíticos , Fraturas Ósseas , Ácido Tranexâmico , Administração Tópica , Perda Sanguínea Cirúrgica/prevenção & controle , Humanos , Hemorragia Pós-Operatória/prevenção & controle , Cloreto de Sódio
11.
J Foot Ankle Surg ; 60(1): 194-198, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33162293

RESUMO

A 31-year-old male was diagnosed with osteoblastic osteosarcoma of the talus. Limb-salvage surgery for talar osteosarcoma was performed by replacing the intact talus with a 3D-printed talar prosthesis made from medical-grade titanium. The prosthesis had 3 tunnels for simulating the ligaments around the talus. At the last follow-up, the functional and clinical outcomes were excellent. Our patient achieved 93% restoration of the Musculoskeletal Tumor Society functional score as well as a Toronto Extremity Salvage Score of 93 points, and there was no local recurrence or distant metastasis. A 3D-printed talar prosthesis showed excellent functional and clinical outcomes for a patient with osteosarcoma of the talus. A 3D-printed implant is a feasible option for patients with osteosarcoma of the foot.


Assuntos
Neoplasias Ósseas , Osteossarcoma , Tálus , Adulto , Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Ósseas/cirurgia , Humanos , Masculino , Recidiva Local de Neoplasia , Osteossarcoma/diagnóstico por imagem , Osteossarcoma/cirurgia , Impressão Tridimensional , Tálus/diagnóstico por imagem , Tálus/cirurgia
12.
Calcif Tissue Int ; 106(5): 518-532, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32189040

RESUMO

Humanin (HN), a mitochondrial derived peptide, plays cyto-protective role under various stress. In this study, we aimed to investigate the effects of HNGF6A, an analogue of HN, on osteoblast apoptosis and differentiation and the underlying mechanisms. Cell proliferation of murine osteoblastic cell line MC3TC-E1 was examined by CCK8 assay and Edu staining. Cell apoptosis was detected by Annexin V assay under H2O2 treatment. The differentiation of osteoblast was determined by Alizarin red S staining. We also tested the expression of osteoblast phenotype related protein by real-time PCR and Western blot. The interaction between Circ_0001843 and miR-214, miR-214 and TAFA5 was examined by luciferase report assay. Circ_0001843 was inhibited by siRNA and miR-214 was suppressed by miR-214 inhibitor to determine the effects of Circ_0001843 and miR-214 on cell proliferation, apoptosis, and differentiation. HNGF6A, an analogue of HN, exerted cyto-protection and osteogenesis-promotion in MC3T3-E1 cells. The expression of osteoblast phenotype related protein was significantly induced by HNGF6A. Additionally, HNGF6A treatment decreased Circ_0001843 and increased miR-214 levels, as well as inhibited the phosphorylation of p38 and JNK. We further found that Circ_0001843 directly bound with miR-214, which in turn inhibited the phosphorylation of p38 and JNK. Furthermore, both Circ_0001843 overexpression and miR-214 knockdown significantly decreased the cyto-protection and osteogenic promotion of HNGF6A. In summary, our data showed that HNGF6A protected osteoblasts from oxidative stress-induced apoptosis and osteoblast phenotype inhibition by targeting Circ_0001843/miR-214 pathway and the downstream kinases, p38 and JNK.


Assuntos
Apoptose , Peptídeos e Proteínas de Sinalização Intracelular/farmacologia , MicroRNAs , Osteoblastos , Estresse Oxidativo , RNA Circular/metabolismo , Células 3T3 , Animais , Diferenciação Celular , Camundongos , MicroRNAs/metabolismo , Osteoblastos/citologia , Fenótipo
13.
J Foot Ankle Surg ; 59(6): 1275-1278, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32830018

RESUMO

Ankle fractures and their mechanisms of injury can be complex. We report a case of a patient with an uncertain mechanism of injury and an uncommon combination of lower extremity fractures, which fit the criteria for a Lauge-Hansen classification pronation-external-rotation fracture, Maisonneuve fracture, Wagstaffe fracture, and posterior pilon fracture. Plain radiographs and computed tomography scan revealed Chaput tubercle avulsion fractures, an anterior distal fibular fracture fragment, multiple lateral malleolar fractures, a posterior malleolar fracture fragment with proximal displacement, a die-punch fragment between the posterior malleolar fragment and the tibia, a proximal fibular fracture, and possible ankle syndesmotic diastasis. Intraoperative hook test was negative after fixation of the fractures, so syndesmotic fixation was not performed. At 3-month follow-up, plain radiographs showed obvious syndesmotic diastasis. At 1-year follow-up, symptoms persisted and syndesmotic fusion was recommended but declined by the patient. This case demonstrates that both ankle fractures and their mechanisms of injury can be remarkably complex and confusing, posterior pilon fractures can occur along with pronation-external-rotation ankle fractures, syndesmotic fixation should be considered for all patients with Maisonneuve fractures, reliance on the hook test for surgical management decisions may not always be reliable, and there exists a need for a more accurate and reliable intraoperative test to determine the presence of ankle syndesmotic injury.


Assuntos
Fraturas do Tornozelo , Traumatismos do Tornozelo , Fraturas da Tíbia , Fraturas do Tornozelo/diagnóstico por imagem , Fraturas do Tornozelo/cirurgia , Traumatismos do Tornozelo/diagnóstico por imagem , Traumatismos do Tornozelo/cirurgia , Articulação do Tornozelo/diagnóstico por imagem , Articulação do Tornozelo/cirurgia , Fixação Interna de Fraturas , Humanos , Estudos Retrospectivos , Fraturas da Tíbia/diagnóstico por imagem , Fraturas da Tíbia/cirurgia
14.
J Cell Biochem ; 119(12): 10351-10357, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30136304

RESUMO

Mice with disruption of Lrrk1 and patients with nonfunctional mutant Lrrk1 exhibit severe osteopetrosis phenotypes because of osteoclast cytoskeletal dysfunction. To understand how Lrrk1 regulates osteoclast function by modulating cytoskeleton rearrangement, we examined the proteins that are differentially phosphorylated in wild-type mice and Lrrk1-deficient osteoclasts by metal affinity purification coupled liquid chromatography/mass spectrometry (LC/MS) analyses. One of the candidates that we identified by LC/MS is L-plastin, an actin bundling protein. We found that phosphorylation of L-plastin at serine (Ser) residues 5 was present in wild-type osteoclasts but not in Lrrk1-deficient cells. Western blot analyses with antibodies specific for Ser5 phosphorylated L-plastin confirmed the reduced L-plastin Ser5 phosphorylation in Lrrk1 knockout (KO) osteoclasts. micro computed tomography (Micro-CT) analyses revealed that the trabecular bone volume of the distal femur was increased by 27% in the 16 to 21-week-old L-plastin KO females as compared with the wild-type control mice. The ratio of bone volume to tissue volume and connectivity density were increased by 44% and 47% (both P < 0.05), respectively, in L-plastin KO mice. Our data suggest that targeted disruption of L-plastin increases trabecular bone volume, and phosphorylation of Ser5 in L-plastin in the Lrrk1 signaling pathway may in part contribute to actin assembly in mature osteoclasts.


Assuntos
Actinas/genética , Glicoproteínas de Membrana/genética , Proteínas dos Microfilamentos/genética , Osteopetrose/genética , Proteínas Serina-Treonina Quinases/genética , Animais , Osso Esponjoso/crescimento & desenvolvimento , Osso Esponjoso/metabolismo , Citoesqueleto/genética , Humanos , Camundongos , Camundongos Knockout , Osteoclastos/metabolismo , Osteoclastos/patologia , Osteopetrose/patologia , Fosforilação/genética , Ligação Proteica , Proteínas Serina-Treonina Quinases/deficiência , Serina/genética , Transdução de Sinais/genética
15.
Cell Physiol Biochem ; 48(1): 293-303, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30016763

RESUMO

BACKGROUND/AIMS: Narrowing of the lumbar spinal canal is a condition called lumbar spinal stenosis (LSS) and is a high-morbidity problem in the elderly. LSS is commonly caused by hypertrophy of the ligamentum flavum (HLF). Previous studies showed that fibrosis of the ligamentum flavum (LF) largely contributed to HLF. However, the underlying pathomechanism remains unclear. Insulin-like growth factor-1 (IGF-1) is known to have an intimate relationship with fibrosis in various tissues. Nevertheless, currently, there are few studies regarding IGF-1 in HLF. In this study, we investigated the role of IGF-1 in HLF and its potential molecular mechanism of action. METHODS: First, the IGF-1, phosphorylation of IGF-1 receptor (pIGF-1R), phosphorylation of AKT (pAKT), phosphorylation of S6(pS6), collagen I and collagen III expression levels were examined via immunohistochemistry and Western blotting in LF tissues from patients with LSS or Non-LSS. Second, primary LF cells were isolated from adults with a normal LF thickness and were cultured with different concentrations of IGF-1 with or without NVP-AEW541/rapamycin. RESULTS: The results showed that IGF-1, pIGF-1R, pAKT, pS6, collagen I and collagen III protein expression in the LSS group was significantly higher than that in the Non-LSS group. Meanwhile, pIGF-1R, pAKT, pS6, collagen I and collagen III protein expression was significantly enhanced in LF cells after IGF-1 exposure, which can be notably blocked by NVP-AEW541. In addition, pS6, collagen I and collagen III protein expression was blocked by rapamycin. CONCLUSIONS: Enhanced IGF-1 promotes the synthesis of collagen I and collagen III via the mTORC1 signaling pathway, which eventually contributes to hypertrophy of the ligamentum flavum.


Assuntos
Hipertrofia/patologia , Fator de Crescimento Insulin-Like I/metabolismo , Ligamento Amarelo/metabolismo , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Transdução de Sinais , Idoso , Estudos de Casos e Controles , Sobrevivência Celular , Colágeno Tipo I/metabolismo , Colágeno Tipo III/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Ligamento Amarelo/citologia , Ligamento Amarelo/diagnóstico por imagem , Masculino , Alvo Mecanístico do Complexo 1 de Rapamicina/antagonistas & inibidores , Pessoa de Meia-Idade , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor IGF Tipo 1/metabolismo , Proteínas Quinases S6 Ribossômicas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia
16.
Am J Physiol Endocrinol Metab ; 311(4): E772-E780, 2016 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-27600824

RESUMO

Leucine-rich repeat kinase-1 (Lrrk1) consists of ankyrin repeats (ANK), leucine-rich repeats (LRR), a GTPase-like domain of Roc (ROC), a COR domain, a serine/threonine kinase domain (KD), and WD40 repeats (WD40). Previous studies have revealed that knockout (KO) of Lrrk1 in mice causes severe osteopetrosis, and a human mutation of Lrrk1 leads to osteosclerotic metaphysial dysplasia. The molecular mechanism by which Lrrk1 regulates osteoclast function is unknown. In this study, we generated a series of Lrrk1 mutants and evaluated their ability to rescue defective bone resorption in Lrrk1-deficient osteoclasts by use of pit formation assays. Overexpression of Lrrk1 or LRR-truncated Lrrk1, but not ANK-truncated Lrrk1, WD40-truncated Lrrk1, Lrrk1-KD, or K651A mutant Lrrk1, rescued bone resorption function of Lrrk1 KO osteoclasts. We next examined whether RAC1/Cdc42 small GTPases are direct substrates of Lrrk1 in osteoclasts. Western blot and pull-down assays revealed that Lrrk1 deficiency in osteoclasts resulted in reduced phosphorylation and activation of RAC1/Cdc42. In vitro kinase assays confirmed that recombinant Lrrk1 phosphorylated RAC1-GST protein, and immunoprecipitation showed that the interaction of Lrrk1 with RAC1 occurred within 10 min after RANKL treatment. Overexpression of constitutively active Q61L RAC1 partially rescued the resorptive function of Lrrk1-deficient osteoclasts. Furthermore, lack of Lrrk1 in osteoclasts led to reduced autophosphorylation of p21 protein-activated kinase-1 at Ser144, catalyzed by RAC1/Cdc42 binding and activation. Our data indicate that Lrrk1 regulates osteoclast function by directly modulating phosphorylation and activation of small GTPase RAC1/Cdc42 and that its function depends on ANK, ROC, WD40, and kinase domains.


Assuntos
Proteínas Ativadoras de GTPase/metabolismo , Neuropeptídeos/metabolismo , Osteoclastos/fisiologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo , Animais , Repetição de Anquirina , Reabsorção Óssea/genética , Masculino , Camundongos , Camundongos Knockout , Mutação/genética , Fosforilação , Proteínas Serina-Treonina Quinases/deficiência , Ligante RANK/farmacologia , Células RAW 264.7
17.
Orthop Surg ; 16(3): 523-531, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38272834

RESUMO

There has been increasing application of autologous costal chondral/osteochondral transplantation (ACCT/ACOT) and costa-derived chondrocyte implantation (ACCI) for articular cartilage repair over the past three decades. This review presents the major evidence on the properties of costal cartilage and bone and their qualifications as grafts for articular cartilage repair, the major clinical applications, and the risks and strategies for costal chondral/osteochondral graft(s) harvest. First, costal cartilage has many specific properties that help restore the articular surface. Costa, which can provide abundant cartilage and cylindrical corticocancellous bone, preserves permanent chondrocyte and is the largest source of hyaline cartilage. Second, in the past three decades, autologous costal cartilage-derived grafts, including cartilage, osteochondral graft(s), and chondrocyte, have expanded their indications in trauma and orthopaedic therapy from small to large joints, from the upper to lower limbs, and from non-weight-bearing to weight-bearing joints. Third, the rate of donor-site complications of ACCT or ACOT is low, acceptable, and controllable, and some skills and accumulated experience can help reduce the risks of ACCT and ACOT. Costal cartilage-derived autografting is a promising technique and could be an ideal option for articular chondral lesions with or without subchondral cysts. More high-quality clinical studies are urgently needed to help us further understand the clinical value of such technologies.


Assuntos
Cartilagem Articular , Cartilagem Costal , Procedimentos Ortopédicos , Humanos , Cartilagem Articular/cirurgia , Cartilagem Articular/lesões , Condrócitos/transplante , Transplante Autólogo
18.
Sci Adv ; 10(6): eadk6722, 2024 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-38324693

RESUMO

Reconstructing extensive cranial defects represents a persistent clinical challenge. Here, we reported a hybrid three-dimensional (3D) printed scaffold with modification of QK peptide and KP peptide for effectively promoting endogenous cranial bone regeneration. The hybrid 3D printed scaffold consists of vertically aligned cryogel fibers that guide and promote cell penetration into the defect area in the early stages of bone repair. Then, the conjugated QK peptide and KP peptide further regulate the function of the recruited cells to promote vascularization and osteogenic differentiation in the defect area. The regenerated bone volume and surface coverage of the dual peptide-modified hybrid scaffold were significantly higher than the positive control group. In addition, the dual peptide-modified hybrid scaffold demonstrated sustained enhancement of bone regeneration and avoidance of bone resorption compared to the collagen sponge group. We expect that the design of dual peptide-modified hybrid scaffold will provide a promising strategy for bone regeneration.


Assuntos
Osteogênese , Alicerces Teciduais , Criogéis , Regeneração Óssea/fisiologia , Peptídeos , Impressão Tridimensional
19.
Foot Ankle Int ; 45(4): 383-392, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38445607

RESUMO

BACKGROUND: Whether arthroscopic microfracture is effective in treating cystic osteochondral lesions of the talus (OLTs) remains controversial. In this study, outcome parameters in patients with small and shallow subchondral cysts are compared to patients without cysts with the hypothesis that equivalent outcomes may be found after primary microfracture treatment. METHODS: From 2018 to 2021, all 50 OLTs treated with arthroscopic microfracture in the authors' hospital were retrospectively reviewed for eligibility. Single unilateral symptomatic lesions were included and divided into the cyst and noncyst groups, whereas kissing lesions and arthritic lesions were excluded. Numeric rating scale (NRS) scores, American Orthopaedic Foot & Ankle Society (AOFAS) ankle-hindfoot scores, Tegner activity level scores, Foot and Ankle Ability Measure (FAAM) scores, and magnetic resonance (MR) imaging results were used to describe outcomes. RESULTS: A total of 35 patients were included, 16 in the cyst group and 19 in the noncyst group. The patient characteristics were similar between the 2 groups (P > .05). In the cyst group the average cysts depth was 5.0 ± 1.3 mm. After a mean follow-up duration of 36.2 ± 10.2 months, no significant differences were found between the 2 groups in NRS, AOFAS, FAAM, or Tegner score improvement (P > .05). Three patients (19%) in the cyst group had no NRS score improvement. CONCLUSION: OLTs with small and shallow subchondral cysts can be treated with arthroscopic microfracture and achieve similar outcomes as noncystic lesions. A few cystic lesions may not respond to microfracture treatment. LEVEL OF EVIDENCE: Level III, retrospective comparative study.


Assuntos
Artroscopia , Cistos Ósseos , Tálus , Humanos , Tálus/cirurgia , Tálus/lesões , Estudos Retrospectivos , Feminino , Artroscopia/métodos , Adulto , Masculino , Cistos Ósseos/cirurgia , Cistos Ósseos/diagnóstico por imagem , Pessoa de Meia-Idade , Imageamento por Ressonância Magnética , Artroplastia Subcondral/métodos , Cartilagem Articular/cirurgia , Cartilagem Articular/lesões
20.
Orthop Surg ; 2024 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-39291799

RESUMO

OBJECTIVE: Few studies have explored the relationship between the pathological characteristics of hallux valgus and surgical outcomes. The aim of our study was to report the influence of pathological characteristics such as the tibial sesamoid position (TSP) and first metatarsal pronation on postoperative functional scores and patient satisfaction with hallux valgus surgery. METHODS: From June 2017 to December 2022, a retrospective analysis was conducted on patients who underwent hallux valgus surgery at our hospital. Anteroposterior (AP) x-ray parameters (hallux valgus angle [HVA], intermetatarsal angle [IMA], distal metatarsal articular angle [DMAA], TSP, first metatarsal pronation, and first metatarsophalangeal joint dislocation) (preoperative weight-bearing, immediate postoperative non-weight-bearing, and early postoperative weight-bearing), visual analog scale (VAS) scores, American Orthopaedic Foot and Ankle Society (AOFAS) forefoot scores, SAFE-Q self-administered foot evaluation questionnaire (excluding the Sports Activity subscale), complications, and patient satisfaction were used to describe the outcomes. Correlation analysis and multiple linear regression models were used to identify factors influencing postoperative functional scores and patient satisfaction after hallux valgus surgery. RESULTS: Eighty-one patients (92 feet) whose early radiographic parameters and latest follow-up scores improved significantly (p < 0.01) were included in the present study. The overall complication rate was 27.2%, with recurrence being the most common complication (20.7%). Functional scores and patient satisfaction were associated with age; preoperative, immediate postoperative, and early postoperative HVA; and immediate postoperative IMA (p < 0.05), but not with TSP, first metatarsal pronation, DMAA, callosities, toe deformities, or first metatarsophalangeal joint dislocation (p > 0.05). The R2 values of the multiple linear regression models predicting postoperative functional scores and patient satisfaction ranged from 0.042 to 0.351. CONCLUSION: TSP and first metatarsal pronation were unrelated to postoperative functional scores or patient satisfaction. This finding enhances surgeons' understanding of the functional prognosis of hallux valgus surgery, particularly aiding in explaining the condition and assessing surgical outcomes.

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