Osteoarthritis, the most common degenerative joint disease, significantly impairs life quality and labor capability of patients. Synovial inflammation, initiated by HMGB1 (High mobility group box 1)-induced activation of macrophage, precedes other pathological changes. As an upstream regulator of NF-κB (nuclear factor-kappa B) and MAPK (mitogen-activated protein kinase) signaling pathway, TAK1 (TGF-β activated kinase 1) participates in macrophage activation, while its function in osteoarthritis remains unveiled. This study aims to investigate the role of TAK1 in the pathogenesis of osteoarthritis via both in vitro and in vivo approaches.

We performed immunohistochemical staining for TAK1 in synovial tissue, both in osteoarthritis patients and healthy control. Besides, immunofluorescence staining for F4/80 as macrophage marker and TAK1 were conducted as well. TAK1 expression was examined in RAW264.7 macrophages stimulated by HMGB1 via qPCR (Quantitative polymerase chain reaction) and Western blotting, and the effect of TAK1 inhibitor (5z-7 oxozeaenol) on TNF-α production was evaluated by immunofluorescence staining. Further, we explored the influence of intra-articular shRNA (short hairpin RNA) targeting TAK1 on collagenase-induced osteoarthritis in mice.

Immunohistochemical staining confirmed significant elevation of TAK1 in osteoarthritic synovium, and immunofluorescence staining suggested macrophages as predominant residence of TAK1. In HMGB1-stimulated RAW264.7 macrophages, TAK1 expression was up-regulated both in mRNA and protein level. Besides, TAK1 inhibitor significantly impairs the production of TNF-α by macrophages upon HMGB1 stimulation. Moreover, intra-articular injection of lentivirus loaded with shRNA targeting TAK1 (sh-TAK1) reduced peri-articular osteophyte formation in collagenase-induced osteoarthritis in mice.

TAK1 exerts a potent role in the pathogenesis of osteoarthritis by mediating the activation of macrophages.

For patients who took joint replacement, one of the complications, aseptic joint loosening, could cause a high risk of revision surgery. Studies have shown that MSCs have the ability of homing and differentiating, and also have highly effective immune regulation and anti-inflammatory effects. However, few studies had focused on the stem cells in preventing the occurrence and development of aseptic loosening. In this research, we aimed to clarify whether human umbilical cord mesenchymal stem cells could inhibited the aseptic joint loosening caused by wear particles.

A Cranial osteolysis mice model was established on mice to examine the effect of hUC-MSCs on the Titanium particles injection area through micro-CT. The amount of stem cells injected was 2 × 10 5 cells. One week later, the mouse Cranial were obtained for micro-CT scan, and then stained with HE analysis immunohistochemical analysis of TNF-α, CD68, CCL3 and Il-1β.

All mice were free of fever and other adverse reactions, and there was no death occurred. Titanium particles caused the osteolysis at the mice cranial, while local injection of hUC-MSCs did inhibit the cranial osteolysis, with a lower BV/TV and a higher porosity. Immunohistochemical results suggested that the expression of TNF-α, CD68, CCL3 and Il-1β in the cranial in Titanium particles mice increased significantly, but was significantly reduced in mice injected with hUC-MSCs. The inhibited CD68 expression indicated that the number of macrophage was lower, which might be a result of the inhibition of CCL3.

According to the studies above, HUC-MSCs treatment of mouse cranial osteolysis model can significantly reduce osteolysis, inhibit macrophage recruitment, alleviate inflammatory response, without causing adverse reactions. It may become a promising treatment of aseptic joint loosening.

Based on Ilizarov's law of tension-stress principle, distraction histogenesis technique has been widely applied in orthopaedic surgery for decades. Derived from this technique, cranial bone transport technique was mainly used for treating cranial deformities and calvarial defects. Recent studies reported that there are dense short vascular connections between skull marrow and meninges for immune cells trafficking, highlighting complex and tight association between skull and brain. Alzheimer's disease (AD) is a progressive neurodegenerative disease and the most common cause of dementia without effective therapy. Meningeal lymphatics have been recognized as an important mediator in neurological diseases. The augmentation of meningeal lymphatic drainage might be a promising therapeutic target for AD. Our proof-of-concept study has indicated that cranial bone transport can promote ischemic stroke recovery via modulating meningeal lymphatic drainage function, providing a rationale for treating AD using cranial bone maneuver (CBM). This study aims to investigate the effects of CBM on AD and to further explore the potential mechanisms.

Transgenic 5xFAD mice model was used in this study. After osteotomy, a bone flap was used to perform CBM without damaging the dura. Open filed test, novel object recognition test and Barn's maze test were used to evaluate neurological functions of 5xFAD mice after CBM treatment. Congo red and immunofluorescence staining were used to evaluate amyloid depositions and Aβ plaques in different brain regions. Lymphangiogenesis and the level of VEGF-C were examined after CBM treatment. OVA-A647 was intra-cisterna-magna injected to evaluate meningeal lymphatic drainage function after CBM treatment.

CBM significantly improved memory functions and reduced amyloid depositions and Aβ plaques in the hippocampus of 5xFAD mice. A significant increase of meningeal lymphatic vessels in superior sagittal sinus and transverse sinus, and the upregulation of VEGF-C in meninges were observed in 5xFAD mice treated with CBM. Moreover, CBM remarkably enhanced meningeal lymphatic drainage function in 5xFAD mice (n=5-16 mice/group for all studies).

CBM may promote meningeal lymphangiogenesis and lymphatic drainage function through VEGF-C-VEGFR3 pathway, and further reduce amyloid depositions and Aβ plaques and alleviate memory deficits in AD.

Aseptic inflammation is the main factor causing aseptic loosening of artificial joints. Studies have shown that inflammatory cells can activate STING (stimulator of interferon genes, STING) after being stressed. This study aims to explore the specific mechanism of STING in aseptic loosening of artificial joints, and provide new strategies for disease prevention.

Titanium particles with a diameter of 1.2-10 μm were prepared to stimulate macrophages (RAW 264.7) to simulate the periprosthetic microenvironment. A lentiviral vector targeting the STING gene was designed and transfected into macrophages to construct a cell line targeting STING knockdown. The expression and secretion levels of TNF-α were detected by qPCR and ELISA, the activation levels of inflammatory pathways (NF-κB, IRF3, etc.) were detected by western blot, and the nucleus translocation of P65 and IRF3 was observed by cellular immunofluorescence.

After titanium particles stimulated macrophages, qPCR and ELISA showed that the transcription and secretion levels of TNF-α were significantly increased. Western blot showed that titanium particle stimulation could increase the phosphorylation levels of NF-κB and IRF3 pathways. While knockdown of STING can significantly reduce titanium particle-induced TNF production, attenuate the activation levels of NF-κB and IRF3 pathways as well as the nucleus translocation of P65 and IRF3.

Conclusions: STING positively regulates the level of inflammation in macrophages induced by titanium particles, and targeted inhibition of STING can reduce inflammation, which may delay the progression of aseptic loosening of artificial joints.

Osteoarthritis (OA), the most prevalent chronic joint disease, represents a relevant social and economic burden worldwide. Human umbilical cord mesenchymal stem cells (HUCMSCs) have been used for injection into the joint cavity to treat OA. The aim of this article is to clarify whether Huc-MSCs derived exosomes could inhibit the progression of OA and the mechanism in this process.

A rabbit OA model was established by the transection of the anterior cruciate ligament. The effects of HUCMSCs or exosomes derived from HUCMSCs on repairing articular cartilage of knee osteoarthritis was examined by micro-CT. Immunohistochemical experiments were used to confirm the expression of relevant inflammatory molecules in OA. In vitro experiments, Transwell assay was used to assess the migration of macrophages induced by TNF-a.

Results showed that a large number of macrophages migrated in arthcular cavity in OA model in vivo, while local injection of HUCMSCs and exosomes did repair the articular cartilage. Immunohistochemical results suggested that the expression of CCL2 and CD68 in the OA rabbit model increased significantly, but was significantly reduced by HUCMSCs or exosomes. Transwell assay showed that both HUCMSCs and exosomes can effectively inhibit the migration of macrophage.

In conclusion, the exosomes derived by HUCMSCs might might rescue cartilage defects in rabbit through its anti-inflammatory effects through inhibiting CCL2.

As peri-prosthetic aseptic loosening is one of the main causes of implant failure, inhibiting wear particles induced macrophages inflammation is considered as a promising therapy for AL to expand the lifespan of implant. Here, we aim at exploring the role of p110δ, a member of class IA PI3K family, and Krüppel-like factor 4 (KLF4) in titanium particles (TiPs) induced macrophages-inflammation and osteolysis.

Firstly, IC87114, the inhibitor of p110δ and siRNA targeting p110δ were applied and experiments including ELISA and immunofluorescence assay were conducted to explore the role of p110δ. Sequentially, KLF4 was predicted as the transcription factor of p110δ and the relation was confirmed by dual luciferase reporter assay. Next, assays including RT-PCR, western blotting and flow cytometry were performed to ensure the specific role of KLF4. Finally, TiPs-induced mice cranial osteolysis model was established, and micro-CT scanning and immunohistochemistry assay were performed to reveal the role of p110δ and KLF4 in vivo.

Here, we found that p110δ was upregulated in TiPs-stimulated macrophages. The inhibition of p110δ or knockdown of p110δ could significantly dampen the TiPs-induced secretion of TNFα and IL-6. Further mechanistic studies confirmed that p110δ was responsible for TNFα and IL-6 trafficking out of Golgi complex without affecting their expression in TiPs-treated macrophages. Additionally, we explored the upstream regulators and confirmed that Krüppel-like factor 4 (KLF4) was the transcription repressor of p110δ. Apart from that, KLF4, targeted by miR-92a, could also attenuate TiPs-induced inflammation by mediating NF-κB pathway and M1/M2 polarization. By the establishment of TiPs-induced mice cranial osteolysis model, we found that KLF4 knockdown exacerbated TiPs-induced osteolysis which was strikingly ameliorated by knockdown of p110δ.

In summary, our study suggests the key role of miR-92a/KLF4/p110δ signal in TiPs-induced macrophages inflammation and osteolysis.

Aims

The decrease in the number of satellite cells (SCs), contributing to myofibre formation and reconstitution, and their proliferative capacity, leads to muscle loss, a condition known as sarcopenia. Resistance training can prevent muscle loss; however, the underlying mechanisms of resistance training effects on SCs are not well understood. We therefore conducted a comprehensive transcriptome analysis of SCs in a mouse model.

Aims

Osteoarthritis (OA) is prevalent among the elderly and incurable. Intra-articular parathyroid hormone (PTH) ameliorated OA in papain-induced and anterior cruciate ligament transection-induced OA models; therefore, we hypothesized that PTH improved OA in a preclinical age-related OA model.

Aims

Ageing-related incompetence becomes a major hurdle for the clinical translation of adult stem cells in the treatment of osteoarthritis (OA). This study aims to investigate the effect of stepwise preconditioning on cellular behaviours in human mesenchymal stem cells (hMSCs) from ageing patients, and to verify their therapeutic effect in an OA animal model.

Aims

Many biomechanical studies have shown that the weakest biomechanical point of a rotator cuff repair is the suture-tendon interface at the medial row. We developed a novel double rip-stop (DRS) technique to enhance the strength at the medial row for rotator cuff repair. The objective of this study was to evaluate the biomechanical properties of the DRS technique with the conventional suture-bridge (SB) technique and to evaluate the biomechanical performance of the DRS technique with medial row knots.

Objectives

This study looked to analyse the expression levels of microRNA-140-3p and microRNA-140-5p in synovial fluid, and their correlations to the severity of disease regarding knee osteoarthritis (OA).

Objectives

Distraction osteogenesis (DO) mobilises bone regenerative potential and avoids the complications of other treatments such as bone graft. The major disadvantage of DO is the length of time required for bone consolidation. Mesenchymal stem cells (MSCs) have been used to promote bone formation with some good results.

Objective

The development of surgical site infection in the early weeks following open reduction and internal fixation (ORIF) is a challenging problem. There are no evidence-based guidelines to direct the number of surgical debridements prior to definitive wound closure. The purpose of this study was to assess the success of infection resolution, and to identify risk factors for failure, in post-operative infections treated with a single debridement and primary wound closure.

Resection of the distal femur to properly fit the prosthetic component is a crucial step for prosthesis alignment during TKA. In this study, we development a new integrative(Five-In-One, FIO) femur resection method, which performs distal femoral resection in one procedure instead of the standard five cutting steps. The accuracy and operating time are comparable to the conventional cutting methods using foam bone model experimentation and in 9 patients.

In vitro comparison: Uniformly-sized foam bone femur models were used in this study. New five-in-one cutting devices and conventional cutting devices of the same prosthetic size of #44 were installed and resection of the distal femur by five experienced orthopaedic surgeons. Each surgeon performed five cases with each device. Then a femoral prosthesis (#44) was installed on each cut femur mode and anterior-posterior and lateral X-ray radiographs were taken. The angles facet length and distance between anterior and posterior oblique facets were then measured with goniometer and vernier calliper. The corresponding angles from a standard femoral prosthesis (#44) were also measured. The angular difference between the resection femur and prosthesis was calculated and named Angular Deviation. The valgus angle and anteversion angle were measured on anteriorposterior and lateral X-ray radiographs respectively. The mean value from the five measurements obtained from each surgeon using the same cutting method was used for the comparison of the modified and standard resection model. The operating time of each cutting procedure was recorded. students’t-tests were used for the statistical analysis.

In vivo following up: 9 patients with use of the five-in-one cutting instrument and the Deluxe prosthesis have been evaluated during operating, and followed up for at least one year. Operating time were recorded and HSS clinical and functional scoring systems before the surgery, three months and one year after surgery.

The angular deviation of the new FIO Cutting Device was significantly less than the conventional device in all four anatomic measurements (p < 0.05). The distance deviation in the FIO group was also significantly less in the FIO group compared to the conventional procedure (p < 0.05). The average valgus angle and anteversion angle of the five-in-one cutting device which were measured on anterior-posterior and lateral X-ray radiographs respectively were 6.86° and 3.02° respectively. They were not significantly different when compared with the data of the conventional cutting device, which were 6.56o and 3.06o respectively. The mean of the cutting time of the five-in-one device was 9.70 minutes, which was significantly less than the conventional cutting device which was 21.84 minutes averaged (p < 0.05).

Our data demonstrated that the angular accuracy of the distal femoral resection with the newly Five-in-one technique was greatly improved compared to the conventional cutting method. With the use of the new technique, operative time was also shortened over two folds compared to the conventional method. We conclude that the new five-in-one cutting device is more accurate and shorten operating time compared with the conventional device in the vitro study.

The mean HSS score before surgery was 48.69, 84.7 three months after surgery, and 85.6 at one year after surgery. The survivorship was 100% of patients.

Introduction/Background: Diabetes Mellitus (DM) is a disease affecting over 21 million Americans resulting in numerous systemic effects, one of which is impaired bone healing. This study was designed to determine the osteoinductive capacity of MSC augmentation in allograft incorporation within a critical size femoral defect model in rats with DM.

Materials/Methods: A 5mm critical size defect was created in the right femur of 40 male DM BB Wistar. Groups: DM/Allograft(All)+DBM and Non-DM/All+DBM: In two groups, the defect was filled with an allograft from a normal non-DM donor rat filled with approximately 0.05mm3 of DBM. DM/All+DBM/MSCs: In the experimental group, the defect was filled with All+DBM, and loaded with 10×106 MSC/mL. Histomorphometry: Animals were sacrificed at 4 and 8 weeks post-surgery, the femurs were processed for undecalcified histomorphometry, and seven areas of interest were measured.

Results: At 4 and 8 weeks the average bone formation within the defect and total bone formation in the DM/All+DBM/MSC group and at 8 weeks the average total bone formation in the Non-DM/All+DBM group was significantly increased compared to the DM/All+DBM group. No significance was found comparing the Non-DM/All+DBM and DM/All+DBM/MSC groups.

Discussion/Conclusions: This study reveals decreased amount of new bone formation in DM animals compared to Non-DM animals, showing the detrimental effects of DM upon allograft incorporation in a critical size defect. MSC augmentation resulted in new bone formation in DM animals similar to Non-DM animals, suggesting a potential role for MSC as an advjuvant during the process of allograft incorporation.

Introduction/Background: This study was designed to determine the osteoinductive capacity of rhBMP-2 in a non-critical size femoral defect in normal rats and rats with diabetes mellitus (DM). It was hypothesized that DM would result in impaired bone regeneration in the femoral non-critical size defects due to reduced bone formation and local delivery of rhBMP-2 would accelerate non-DM healing and normalize impaired bone healing in DM rats to the levels of Non-DM bone healing.

Materials/Methods: A total of 80 BB Wistar rats were used in the project. A 3mm defect was created during surgery and stabilized with a polyimide plate and either a 0.05cc/11μg dose of rhBMP-2 or buffer in a collagen sponge was implanted into the defect. Microradiographs were taken on the day of sacrifice and processing of samples for PECAM-1 immunohistochemistry, histomorphometry, and mechanical testing was performed.

Results: Both Non-DM and DM groups treated with rhBMP-2 demonstrated significantly higher radiographic scoring, total new bone formation, BV quantification, and mechanical testing parameters compared to those treated with buffer at all timepoints with no significance noted between Non-DM and DM groups treated with rhBMP-2.

Discussion/Conclusions: This study reveals decreased amount of new bone formation in DM animals compared to Non-DM animals, showing the detrimental effects of DM upon bone healing. A single application of rhBMP-2 resulted in new bone formation in DM animals similar to Non-DM animals, suggesting a critical role for rhBMP-2 in ameliorating the deleterious effects of DM on bone regeneration and formation.

Besides these groups 15 more DM rats were used for PECAM-1 staining for angiogenesis (7 with 1 loss at a 3 week time point) and mechanical testing (8 at a 9 week time point).

In femoral locked nailing, the distal locking screws is vulnerable to mechanical failure. The stress on these screws is substantially affected by fitness of the nail in the medullary canal. In this study a closed form mathematical model based on elastic column-beam theory is developed to investigate how the nail-cortical contact which is simulated by a linear elastic foundation affects the stress of the distal locking screws. The model is comprised of a construct of a fractured femur with an intramedullay locked nail loaded by an eccentric vertical load. The stress of the locking screw is analysed as a function of the depth of the locking screw in the distal fragment under two situations: with or without nail-cortical contact in the distal fragment. In situation with nail-cortical contact, the screw stress is decreased as the length of nail-cortical contact and the depth of the screw in the distal fragment increases, but this stress contrarily increased when the nail is inserted beyond a certa

Background: Reconstruction nail had developed since 1985. General indications reviewed from literatures are 1. Ipsilateral femoral neck-shaft fracture (nondisplaced), 2. Russell-Type IB subtrochanteric fracturtes (intact piriformis fossa, fractured less trochanter). Many authors did not recommend that application of reconstruction nail in displaced ipsilateral femoral neck-shaft fractures. The reason is that unpredictable femoral neck-shaft reduction and over-distraction of shaft fracture. We developed one new method for overcoming such technical puzzle to achieve one-step reduction for displaced ipsilateral femoral neck-shaft fractures.

Material: There are 24 consecutive cases were treated by reconstruction nail by the same operator in Chi-Mei Foundation Hospital from February, 1999 to June, 2000. Five of them were diagnosed as displaced ipsilateral femoral neck-shaft fractures and treating new surgical technique in reconstruction nailing. Initial radiographic assessment revealed displaced neck fracture can be classified as Garden III, the fracture morphology is vertical (Pauwell III). Average age of these five patients is 37.6 y/o. The sex distribution is M:F=3:2

Method: Provisional proximal fixation of femur is mandatory. First, we use two 5.0mm drillpit transfixed trochanter region after assure of femoral anteversion. Second, release of traction and distal locking for reduction and fixation of shaft fracture part. Third, remove application handle and use Internal rotation or other remote maneuver for restoration of neck-shaft angle. Finally, complete drilling through neck and sequent proximal cephalomedullary locking was performed by free-hand method.

Result: Initial reduction result was acceptable. There was no significant coxa-varus or coxa-brevis. Two of them had removed of implants and clinical result was satisfied. No avascular necrosis was noted in our following up.

Discussion: How to treat displaced ipsilateral femoral neck-shaft fractures in one-step was obstacle in our orthopedic practice. Abandonment of reconstruction nail just due to technique demanding purpose is very pity. We developed such technique to make patient with displaced ipsilateral femoral neck-shaft fractures treat by closed and one-step method and gained more satisfaction.