Astragaloside IV Protects Against IL-1ß-Induced Chondrocyte Damage via Activating Autophagy.

BACKGROUND: Osteoarthritis (OA) is a chronic inflammatory condition that affects the articular cartilage. Astragaloside IV (AS-IV) constitutes the primary active component of the Chinese herbal medicine Huangqi (Radix Astragali Mongolici). AS-IV demonstrates anti-inflammatory and anti-apoptotic attributes, exhibiting therapeutic potential across various inflammatory and apoptosis-related disorders. Nevertheless, its pharmaceutical effects in OA are yet to be fully defined. OBJECTIVES: This study aimed to investigate the protective impact of AS-IV on rat chondrocytes treated with IL-1ß and ascertain whether autophagy plays a role in this effect. METHODS: Chondrocytes were isolated and cultivated from the knee joints of neonatal SD mice. The study included the blank control group, the model group, and the AS-IV concentration gradient group (50, 100, 200 µmol/L) to intervene with chondrocytes. The MTT assay was employed to assess cell viability at varying culture periods, enabling the determination of suitable concentration and duration. Subsequently, chondrocytes were treated with the optimal AS-IV concentration and divided into three groups: the model group replicated IL-1ß-induced inflammatory chondrocyte injury, the AS-IV group received a co-culture of AS-IV and IL-1ß, and a blank control group was established. Changes in cell morphology and structure were observed using ghost pen cyclic peptide staining. ELISA was used to measure TNF-α and GAG levels in cell supernatants. RT-qPCR assessed p62 and LC3 mRNA expression, while Western Blot evaluated p62 and LC3Ⅱ/Ⅰ protein expression. RESULTS: AS-IV promoted chondrocyte proliferation and concurrently inhibited cell apoptosis. An optimal AS-IV dose of 200 µmol/L and a suitable reaction time of 48 h were identified. Ghost pen cyclic peptide staining indicated that the model group's cytoskeleton exhibited fusiform changes with reduced immunofluorescence intensity, as opposed to the blank control group. The AS-IV group displayed more polygonal cytoskeletal morphology with increased immunofluorescence intensity. AS-IV reduced TNF-α levels and elevated GAG levels in the culture supernatant. Additionally, AS-IV lowered p62 mRNA and protein expression while increasing LC3 mRNA expression in cultured chondrocytes. CONCLUSION: Our findings suggest that AS-IV mitigates inflammatory chondrocyte injury, safeguarding chondrocytes through a potential autophagy suppression mechanism. These results imply that AS-IV could offer preventive advantages for OA.

Icariin regulates miR-23a-3p-mediated osteogenic differentiation of BMSCs via BMP-2/Smad5/Runx2 and WNT/ß-catenin pathways in osteonecrosis of the femoral head.

Icariin is commonly used for the clinical treatment of osteonecrosis of the femoral head (ONFH). miR-23a-3p plays a vital role in regulating the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). The present study aimed to investigate the roles of icariin and miR-23a-3p in the osteogenic differentiation of BMSCs and an ONFH model. BMSCs were isolated and cultured in vitro using icariin-containing serum at various concentrations, and BMSCs were also transfected with a miR-23a inhibitor. The alkaline phosphatase (ALP) activity and cell viability as well as BMP-2/Smad5/Runx2 and WNT/ß-catenin pathway-related mRNA and protein expression were measured in BMSCs. Additionally, a dual-luciferase reporter assay and pathway inhibitors were used to verify the relationship of icariin treatment/miR-23a and the above pathways. An ONFH rat model was established in vivo, and a 28-day gavage treatment and lentivirus transfection of miR-23a-3p inhibitor were performed. Then, bone biochemical markers (ELISA kits) in serum, femoral head (HE staining and Digital Radiography, DR) and the above pathway-related proteins were detected. Our results revealed that icariin treatment/miR-23a knockdown promoted BMSC viability and osteogenic differentiation as well as increased the mRNA and protein expression of BMP-2, BMP-4, Runx2, p-Smad5, Wnt1 and ß-catenin in BMSCs and ONFH model rats. In addition, icariin treatment/miR-23a knockdown increased bone biochemical markers (ACP-5, BAP, NTXI, CTXI and OC) and improved ONFH in ONFH model rats. In addition, a dual-luciferase reporter assay verified that Runx2 was a direct target of miR-23a-3p. These data indicated that icariin promotes BMSC viability and osteogenic differentiation as well as improves ONFH by decreasing miR-23a-3p levels and regulating the BMP-2/Smad5/Runx2 and WNT/ß-catenin pathways.

Icariin Accelerates Fracture Healing via Activation of the WNT1/ß-catenin Osteogenic Signaling Pathway.

BACKGROUND: Icariin has been shown to enhance bone formation. OBJECTIVE: The present study aimed to investigate whether icariin also promotes bone fracture healing and its mechanisms. METHODS: First, we isolated and cultured rat bone marrow stromal cells (rBMSCs) with icariincontaining serum at various concentrations (0%, 2.5%, 5% and 10%) and then measured alkaline phosphatase (ALP) activity and the expression of Core-binding factor, alpha 1 (Cbfα1), bone morphogenetic protein-2 (BMP-2) and bone morphogenetic protein-4 (BMP-4) in the rBMSCs. Second, we established a model of fracture healing in rats and performed gavage treatment for 20 days. Then, we detected bone biochemical markers (ELISA kits) in the serum, fracture healing (digital radiography, DR), and osteocalcin expression (immunohistochemistry). RESULTS: Icariin treatment increased ALP activity and induced the expression of Cbfα1, BMP-2 and BMP-4 in rBMSCs in a dose-dependent manner. In addition, Icariin increased the serum levels of osteocalcin (OC), bone-specific alkaline phosphatase (BAP), N-terminal telopeptides of type I collagen (NTX-1), C-terminal telopeptide of type I collagen (CTX-1) and tartrate-resistant acid phosphatase 5b (TRACP-5b); promoted osteocalcin secretion at the fracture site; and accelerated fracture healing. CONCLUSION: Icariin can promote the levels of bone-formation markers, accelerate fracture healing, and activate the WNT1/ß-catenin osteogenic signaling pathway.

Profiles of immune cell infiltration and immune-related genes in the tumor microenvironment of osteosarcoma.

This work aimed to investigate tumor-infiltrating immune cells (TIICs) and immune-associated genes in the tumor microenvironment of osteosarcoma. An algorithm known as ESTIMATE was applied for immune score assessment, and osteosarcoma cases were assigned to the high and low immune score groups. Immune-associated genes between these groups were compared, and an optimal immune-related risk model was built by Cox regression analyses. The deconvolution algorithm (referred to as CIBERSORT) was applied to assess 22 TIICs for their amounts in the osteosarcoma microenvironment. Osteosarcoma cases with high immune score had significantly improved outcome (P<0.01). The proportions of naive B cells and M0 macrophages were significantly lower in high immune score tissues compared with the low immune score group (P<0.05), while the amounts of M1 macrophages, M2 macrophages, and resting dendritic cells were significantly higher (P<0.05). Important immune-associated genes were determined to generate a prognostic model by Cox regression analysis. Interestingly, cases with high risk score had poor outcome (P<0.01). The areas under the curve (AUC) for the risk model in predicting 1, 3 and 5-year survival were 0.634, 0.781, and 0.809, respectively. Gene set enrichment analysis suggested immunosuppression in high-risk osteosarcoma patients, in association with poor outcome.

Biocompatiable silk fibroin/carboxymethyl chitosan/strontium substituted hydroxyapatite/cellulose nanocrystal composite scaffolds for bone tissue engineering.

Bone defects arise from trauma, skeletal diseases or tumor resections have become a critical clinical challenge. Biocomposite materials as artificial bone repair materials provide a promising approach for bone regeneration. In this study, we used silk fibroin (SF), carboxymethyl chitosan (CMCS), cellulose nanocrystals (CNCs) and strontium substituted hydroxyapatite (Sr-HAp) to prepare the biocomposite scaffolds of SF/CMCS, SF/CMCS/CNCs, SF/CMCS/CNCs/Sr-HAp. The characterization results showed that all the SF-based scaffolds have a porous sponge-like structure with porosities over 80%. In addition, there was a significant increase in compressive strength of SF/CMCS/Sr-HAp/CNCs scaffold when compared to that of SF/CMCS scaffolds, while maintaining high porosity with lower swelling ratio. All the SF-based scaffolds were non-toxic and had a good hemocompatibility. Comparing to the SF/CMCS scaffold, the scaffolds with addition of Sr-HAp and/or CNCs showed enhanced protein adsorption and ALP activity. In addition, higher expression of osteogenic gene markers such as RUNX2, ALP, OCN, OPN, BSP and COL-1 further substantiated the applicability of SF/CMCS/Sr-HAp/CNCs scaffolds for bone related applications. Hence, this study suggests that SF/CMCS/Sr-HAp/CNCs scaffolds have a potential in non-loading bone repair application.

[Comparing hydroxyapatite coated versus non hydroxyapatite coated femoral stems in primary total hip arthroplasty: a meta analysis of randomized controlled trial].

OBJECTIVE: To evaluate the difference of clinical outcomes and radiological outcomes through meta-analysis on the total hip arthroplasty (THA) between hydroxyapatite(HA) coating and non-HA coating femoral stems. METHODS: We searched the MEDLINE, Embase, Cochrane library and CBM for published randomized controlled trial (RCT) comparing HA coating and non-HA coating femoral stems in primary THA clinical outcomes with Harris hip score and incidence postoperative thigh pain, radiological outcomes with presence of endosteal condensation and radioactive line on the prothesis, heterotopic ossification. Data analysis were performed using RevMan 5.0(the Cochrane Collaboration). RESULTS: Ten studies and 917 hips into our analysis, with 464 hips in HA groups and 453 hips in non-HA groups. The combined results of the meta-analysis indicated there was no statistical differences between the two groups on postoperative Harris hip score(WMD = 3.04, 95%CI:-4.47-10.54, P = 0.43) , there was statistical difference on incidence postoperative thigh pain (RR = 0.56, 95%CI:0.33-0.94, P = 0.03) . There were no significant differences between the two groups on presence of endosteal condensation (RR = 1.01, 95%CI:0.91-1.11, P = 0.91), presence of radioactive line (RR = 0.99, 95%CI:0.88-1.11, P = 0.83) and incidence of heterotopic ossification (RR = 0.97, 95%CI:0.77-1.21, P = 0.77). CONCLUSIONS: There are no clinical and radiological benefits in the use of HA coating femoral stems in Primary THA, there is not enough evidence prove the HA can reduce the incidence postoperative thigh pain.