Prenatal ethanol exposure and changes in fetal neuroendocrine metabolic programming.

Prenatal ethanol exposure (PEE) (mainly through maternal alcohol consumption) has become widespread. However, studies suggest that it can cause intrauterine growth retardation (IUGR) and multi-organ developmental toxicity in offspring, and susceptibility to various chronic diseases (such as neuropsychiatric diseases, metabolic syndrome, and related diseases) in adults. Through ethanol's direct effects and its indirect effects mediated by maternal-derived glucocorticoids, PEE alters epigenetic modifications and organ developmental programming during fetal development, which damages the offspring health and increases susceptibility to various chronic diseases after birth. Ethanol directly leads to the developmental toxicity of multiple tissues and organs in many ways. Regarding maternal-derived glucocorticoid-mediated IUGR, developmental programming, and susceptibility to multiple conditions after birth, ethanol induces programmed changes in the neuroendocrine axes of offspring, such as the hypothalamus-pituitary-adrenal (HPA) and glucocorticoid-insulin-like growth factor 1 (GC-IGF1) axes. In addition, the differences in ethanol metabolic enzymes, placental glucocorticoid barrier function, and the sensitivity to glucocorticoids in various tissues and organs mediate the severity and sex differences in the developmental toxicity of ethanol exposure during pregnancy. Offspring exposed to ethanol during pregnancy have a "thrifty phenotype" in the fetal period, and show "catch-up growth" in the case of abundant nutrition after birth; when encountering adverse environments, these offspring are more likely to develop diseases. Here, we review the developmental toxicity, functional alterations in multiple organs, and neuroendocrine metabolic programming mechanisms induced by PEE based on our research and that of other investigators. This should provide new perspectives for the effective prevention and treatment of ethanol developmental toxicity and the early prevention of related fetal-originated diseases.

Analysis of risk factors for advanced age in patients with frozen shoulder.

OBJECTIVE: To explore the risk factors of elderly patients with frozen shoulder. METHODS: 262 cases of scapulohumeral periarthritis treated in our hospital from January 2020 to December 2020 were analyzed retrospectively. According to the age of patients, patients younger than 60 years old were divided into middle-aged group (101 cases), patients between 60 and 75 years old were divided into old-aged group (91 cases), and patients ≥ 75 years old were divided into old-aged group (70 cases). The general demographic data and clinical data of the three groups were compared. Visual analogue scale (VAS) was used to evaluate the degree of pain. Finally, the dependent variable is set as whether the onset age of scapulohumeral periarthritis patients is advanced. Univariate and multivariate Logistic regression was used to analyze the risk factors of frozen shoulder patients at an advanced age. RESULTS: There were no significant differences in general demographic data, fixed position, hypertension history, smoking history, drinking history, supraspinatus muscular atrophy and physical exercise among the three groups (all P > 0.05). The course of disease, diabetes, surgical treatment, pain degree, operation time, cholecystitis, coronary heart disease, pain degree three months after operation and cervical spondylosis in the elderly group were all higher than those in the middle-aged group and the elderly group, and the differences were statistically significant (all P < 0.05). The course of scapulohumeral periarthritis, the degree of pain and the degree of pain 3 months after operation in the elderly group were higher than those in the middle-aged group, with significant differences (all P < 0.05). Univariate Logistic regression analysis showed that the risk factors of scapulohumeral periarthritis in the elderly included diabetes mellitus (OR = 3.067, 95% CI 1.881-4.587, P < 0.001), operative treatment (OR = 3.076, 95% CI 1.365-6.765, P = 0.006), VAS score (OR = 2.267, 95% CI 1.117-3.887, P = 0.013), operation time (OR = 1.537, 95% CI 1.305-2.579, P < 0.001), cholecystitis (OR = 2.143, 95% CI 1.019-4.876, P = 0.023), coronary heart disease(OR = 3.128, 95% CI 1.428-7.019, P = 0.005), VAS at 3 months after operation (OR = 1.537, 95% CI 0.786-2.635, P = 0.002), and cervical spondylosis(OR = 1.162, 95% CI 1.029-1.321, P = 0.012). Multivariate logistic regression analysis showed that the risk factors for the onset of the disease at advanced age included fatty infiltration (OR = 4.021, 95% CI 2.981-9.682, P < 0.001), surgical treatment (OR = 4.109, 95% CI 1.419-7.832, P = 0.008), VAS score (OR = 3.081, 95% CI 1.042-7.931, P = 0.046) and operation time (OR = 1.537, 95% CI 1.305-2.579, P < 0.001). CONCLUSION: Risk factors of frozen shoulder at advanced age include fat infiltration, surgical treatment, VAS score and surgical time. In clinical practice, we should refer to the above indicators to help patients with early medical intervention and prevent their onset.

Simvastatin promotes rat Achilles tendon-bone interface healing by promoting osteogenesis and chondrogenic differentiation of stem cells.

To investigate the effect and mechanism of simvastatin on cell components of tendon-bone healing interface. The tendon-bone healing model was established by inserting the end of the Achilles tendon into the tibial tunnel on 24 rats, and simvastatin was used locally at the tendon-bone interface. Healing was evaluated at 8 weeks by mechanical testing, micro-CT, and qualitative histology including H&E, Toluidine blue, and immunohistochemical staining. In vitro, bone marrow stromal cells (BMSCs) and tendon-derived mesenchymal stem cells (TDSCs) underwent osteogenic and chondrogenic differentiation respectively by plate co-culture. An analysis was performed on days 7 and 14 of cell differentiation. Biomechanical testing demonstrated a significant increase in maximum stiffness in the simvastatin-treated group. Micro-CT analysis showed that the bone tunnels in the simvastatin group were smaller in diameter and had higher bone density. H&E and Toluidine blue staining demonstrated that tendon-bone healing was significantly greater with better tissue arrangement and more extracellular matrix in the simvastatin-treated group than that in the control group, and immunohistochemical staining showed the expression of VEGF in simvastatin group was significantly higher. Histological staining and RT-PCR confirmed that simvastatin could promote the differentiation of co-cultured BMSCs and TDSCs into osteoblasts and chondroblasts, respectively. The effect of promoting osteogenic differentiation was more tremendous at 14 days, while its effect on promoting chondroblast differentiation was more evident on the 7th day of differentiation. In conclusion, local administration of simvastatin can promote the tendon-bone healing by enhancing neovascularization, chondrogenesis, and osteogenesis in different stages of the tendon-bone healing process.

miR-200b-3p/ERG/PTHrP axis mediates the inhibitory effect of ethanol on the differentiation of fetal cartilage into articular cartilage.

PURPOSE: This study aims to further explore cartilage development in prenatal ethanol exposure (PEE) offspring at different times to explore the specific time points and mechanism of ethanol-induced fetal cartilage dysplasia. METHODS: On gestational day (GD)14, GD17, and GD20, PEE fetal cartilage was evaluated by morphological analysis. RT-qPCR, immunohistochemistry, and immunofluorescence were used to detect the expression of cartilage marker genes and their regulatory factors. Bone marrow mesenchymal stem cells (BMSCs) were used to explore the effect of ethanol on the differentiation of chondrocytes. Additionally, we used inhibitors, overexpression plasmids and a luciferase reporter assay on GD17 chondrocytes to verify the mechanism. RESULTS: PEE significantly reduced cartilage matrix content and the expression of marker genes on GD17 and GD20 but had no effect on GD14. The inhibition of chondrogenic differentiation by PEE mainly occurred on GD14-17. Furthermore, the expression of miR-200b-3p was increased, while that of ERG and PTHrP was markedly reduced in PEE fetal cartilage. In vitro, ethanol (30-120 mM) inhibited the differentiation of BMSCs into chondrocytes in a concentration-dependent manner, accompanied by strong expression of miR-200b-3p and low expression of ERG and PTHrP. Moreover, PTHLH and ERG overexpressed, as well as a miR-200b-3p inhibitor reversed the inhibitory effect of ethanol on the differentiation of fetal chondrocytes. Furthermore, miR-200b-3p could target and negatively regulate ERG. CONCLUSIONS: PEE can significantly inhibit the development of articular cartilage, especially during articular cartilage formation. The mechanism is related to the decreased differentiation of fetal cartilage into articular cartilage mediated by the miR-200b-3p/ERG/PTHrP axis.

Pravastatin ameliorated osteoarthritis susceptibility in male offspring rats induced by prenatal ethanol exposure.

Osteoarthritis (OA) is a disease associated with a disorder of cholesterol metabolism. Our previous studies showed that prenatal ethanol exposure (PEE) caused cholesterol accumulation in articular cartilage and increased the susceptibility to OA in offspring. However, we did not determine whether pravastatin, a cholesterol-lowering agent, could rescue PEE-induced susceptibility to OA. Here, fetal rats were divided into a PEE group and a control group during pregnancy. At postnatal week (PW) 8, sixteen male offspring rats from both groups were injected papain through the articular cavity. Eight of them from each group were treated with pravastatin (20 mg/kg·d) by gavage for four weeks simultaneously. We found that pravastatin ameliorated papain-induced high expression of inflammatory factors [interleukin (IL)-1, IL-6], matrix degradation enzymes [matrix metalloproteinase (MMP)-3, MMP-13], and apoptosis factors (caspase-3 and caspase-8) in the cartilage of the PEE group. Also, pravastatin significantly reduced the content of TCH in the blood and cartilage of the PEE offspring and improved cholesterol efflux pathway. Our in vitro findings further confirmed that pravastatin partially reversed cholesterol-induced inflammation and apoptosis of chondrocytes. In conclusion, pravastatin effectively reduced inflammation and matrix degradation, and thus ameliorate OA susceptibility in articular cartilage by relieving cholesterol accumulation in chondrocyte.

Safety and Effectiveness of Orthopaedic Medical Staff in Providing Support in Combating the COVID-19 Pandemic: A Retrospective Investigation from Wuhan, China.

OBJECTIVE: To evaluate whether it is safe and effective for orthopaedic medical staff to provide support to the work against COVID-19. METHODS: One hundred and twenty-two orthopaedic medical staff from the orthopaedic center of Zhongnan Hospital of Wuhan University were included in this retrospective investigation. A total of 43 surgeons and 69 nurses provided medical support in the treatment of COVID-19 patients from 1 January 2020 to 8 April 2020 in four different hospitals in Wuhan. We collected data on the age, gender, and body temperature of orthopaedic medical staff, as well as the results for their chest CT scans, SARS-CoV-2 RNA, SARS-CoV-2 IgM and SARS-CoV-2 IgG tests, and training and examinations on COVID-19 knowledge. We also collected data on the time span of work, the number of infected staff during the support period, the number of COVID-19 patients the surgeons treated and the cure rate, the performance of the surgeons as assessed by the specialists and patients, and the number of infected staff during the pandemic. RESULTS: Among the 49 surgeons and 73 nurses, 43 surgeons and 69 nurses provided support against COVID-19. A total of 12 surgeons and 11 nurses provided support in the fields of respiration, intensive care, and emergency. A total of 34 surgeons and 58 nurses worked in the designated wards restructured for COVID-19 in the orthopaedic building. The average time span of work for the surgeons and nurses was 14.78 ± 3.64 days and 24.77 ± 7.58 days, respectively. No staff were infected during the support period. Over 1000 patients were received in the fever clinic by orthopaedic surgeons. The overall number of the treated hospitalized patients was 622. Among these patients, 226 cases were mild, 318 were mild to moderate, and 58 were severe or critical. The cure rate was 96.01%, 99.37%, and 52.00% respectively. The performance of the surgeons was scored 87.02 ± 3.17 and 90.69 ± 3.58 by the specialists and the patients, respectively. During the whole pandemic, 3 surgeons and 3 nurses who did not participate in the support work were infected in the early stages. The morbidity of all the orthopaedic staff was 4.92% during the whole pandemic, while no one was infected during the support work. CONCLUSION: Our investigation indicated that although they worked outside their specialty, it was safe and effective for the orthopaedic staff to provide medical support in the work against COVID-19 with adequate precautions and proper training.

Identification of differentially expressed genes in synovial tissue of osteoarthritis based on a more robust integrative analysis method.

OBJECTIVE: This study aimed to identify osteoarthritis (OA) related genes based on microarray data in synovium with a more robust integrative analysis method. METHODS: Four series GSE55457, GSE12021, GSE55235, and GSE55584 (36 OA and 29 normal samples) were downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) of GSE55457, GSE12021, and GSE55235 were identified using the LIMMA package. Overlapping DEGs from the intersection of the three series were detected. Simultaneously, samples in the four series were pooled to identify DEGs with integrated analysis using the Sva package. RESULTS: In total, 74 overlapping DEGs and 242 DEGs by integrating four series were detected. Based on them, 70 common DEGs were used to construct a protein-protein interaction (PPI) network, involving 61 nodes and 206 edges. Also, three gene modules and five hub genes, named JUN, IL6, VEGFA, MYC, and EGR1, were identified. CONCLUSIONS: Seventy DEGs were finally identified with a more robust integrative analysis method. JUN, IL6, VEGFA, MYC, and EGR1 were identified as hub genes in the development of OA. Key Points • 76 overlapping DEGs were detected from the intersection of DEGs in GSE55457, GSE12021, and GSE55235. • 242 DEGs were identified by integrating four series using Sva package. • 72 common DEGs were finally identified based on the overlapping DEGs and the integrated DEGs. • JUN, IL6, VEGFA, MYC, and EGR1 were identified as hub genes in the development of OA.

Morphological study of the vastus medialis oblique in recurrent patellar dislocation based on magnetic resonance images.

BACKGROUND: To investigate the morphological parameters of the vastus medialis obliquus (VMO) muscle and delineate its importance in the maintenance of patellofemoral joint stability. METHODS: The magnetic resonance imaging data of seventy-five knees (fifty-four patients) with recurrent lateral patella dislocation (LPD) and seventy-five knees (seventy patients) without recurrent LPD were retrospectively analysed. Five morphological parameters related to the VMO (elevation in the sagittal plane and coronal plane, craniocaudal extent, muscle-fibre angulation, cross-sectional area ratio) and two patella tilt parameters (patella tilt angle, bisect offset ratio) were measured in MR images. The independent-samples t test or chi-square test was used for statistical comparisons. RESULTS: The mean ages of the patients in the recurrent LPD group and control group were 22.1 ± 9.9 years and 24.0 ± 6.5 years, respectively. Eighteen out of seventy-five (24%) patients MRI showed VMO injuries. Compared with the control group, the patients with recurrent LPD showed significantly higher sagittal VMO elevation (10.4 ± 2.3 mm vs. 4.1 ± 1.9 mm), coronal VMO elevation (15.9 ± 5.7 mm vs. 3.9 ± 3.7 mm), muscle-fibre angulation (35.4 ± 8.0° vs. 27.9 ± 6.3°), patella tilt angle (25.9 ± 10.7° vs. 9.1 ± 5.2°), and bisect offset ratio values (0.9 ± 0.3 vs. 0.5 ± 0.1) and significantly lower craniocaudal extent (13.7 ± 5.3 mm vs. 16.7 ± 5.1 mm) and cross-sectional area ratio values (0.05 ± 0.02 vs. 0.07 ± 0.02). CONCLUSIONS: The results showed that abnormalities in the VMO and patella tilt were clearly present in recurrent LPD patients compared with normal people.

The pronator teres and the flexor carpi radialis interval approach for operative fixation of ulna coronoid process fractures.

BACKGROUND: The optimal approach for the fixation of coronoid process fractures is unknown. We present the advantages and the clinical effect of the pronator teres and the flexor carpi radialis interval approach for the treatment of ulna coronoid process fractures. METHODS: The patients, who had operative treatment of closed ulna coronoid process fracture by the pronator teres and the flexor carpi radialis interval approach between January 2011 to December 2016, were studied retrospectively. Seventeen consecutive patients had received surgical fixation by screws or a mini-plate through the above approach, of whom were 16 males and one female with an average age of 36.7 years (range, 21-58 years). There were 11 cases of type II and 6 cases of type III according to the O'Driscoll classification, of which, 6 patients had combined elbow dislocation, 2 patients showed elbow instability after fixation, and one had another incision to repair the lateral collateral ligament, and received a hinged external fixator. The other patient only received a hinged external fixator for 4 weeks. Mayo Elbow Performance Score (MEPS) was used to assess the function of elbow for each patient at the final follow-up. RESULTS: Mean follow-up was 28.7 months (range, 24-38 months). Fracture union was achieved in each patient; the average time to radiologic union was 14.2 weeks (range, 12-16 weeks). At the final follow-up, the elbow extension degree of the affected side was (3.88±2.96°), reaching 98.1% of the normal side, and the flexion degree was (131.59±4.93°), reaching 98.16% of the normal side. The forearm pronation was (82.94±3.86°), reaching 94.31% of the normal side, and the supination activity was (82.12±3.82°), reaching 93% of the normal side. According to the MEPS, the functional recovery of the injured arm was assessed as excellent in 16 cases, and good in one. None of the patients showed any neurovascular or deep infections and no heterotopic ossification was found. CONCLUSIONS: The pronator teres and the flexor carpi radialis interval approach has the advantages of simplicity, safety, minimal invasion, excellent exposure, and good postoperative function recovery for ulna coronoid process fracture.

Comparative study of the tibial tubercle-trochlear groove distance measured in two ways and tibial tubercle-posterior cruciate ligament distance in patients with patellofemoral instability.

BACKGROUND: A comparative analysis of the strengths and weaknesses of three different methods for radiologic evaluation of patellofemoral instability (PFI). METHODS: Computed tomography (CT) and magnetic resonance imaging (MRI) were performed in 47 patients with or without PFI. The tibial tubercle-trochlear groove (TT-TG) distance was measured by two observers through conventional CT and three-dimensional CT reconstruction (TDR-TT-TG) respectively and the tibial tubercle-posterior cruciate ligament (TT-PCL) distance with MRI. The intraclass correlation coefficient (ICC) was used to evaluate the interobserver reliability. In addition, the differences of three measurements between different patients were compared. The consistency of TT-TG and TDR-TT-TG was analyzed by the Bland-Altman method. RESULTS: The ICCs of three measurements were high between two observers; the results were TT-TG (ICC = 0.852), TDR-TT-TG (ICC = 0.864), and TT-PCL (ICC = 0.758). The values of PFI patients were significantly higher than those of non-PFI patients, and the mean TT-TG, TDR-TT-TG, and TT-PCL distance in patients with PFI were 19.0 ± 3.8 mm, 19.0 ± 3.7 mm, and 25.1 ± 3.6 mm, respectively. There was no statistically significant difference between the TT-TG distance and the TDR-TT-TG distance, we found no significant difference. The Bland-Altman analysis showed that the TDR-TT-TG distance was in good agreement with the TT-TG distance. CONCLUSION: All three methods can be used to assess PFI; the TDR-TT-TG measurement method has superior operability and better interobserver consistency. It may be an alternative method to the conventional TT-TG distance measurement.

Dexamethasone causes calcium deposition and degeneration in human anterior cruciate ligament cells through endoplasmic reticulum stress.

BACKGROUND: Dexamethasone is widely used in the treatment of joint diseases due to its anti-inflammatory properties. However, it can cause serious adverse effects. The anterior cruciate ligament (ACL) is an important stabilizer of the knee joint. However, the effect of dexamethasone treatment on the ACL is unclear. OBJECTIVE: This study aims to explore the effects of dexamethasone on ACL tissues and cells through in vitro and in vivo experiments. RESULTS: In vitro, we found that after treatment with dexamethasone, human ACL cell apoptosis was increased, type I collagen (COL1A1) content was decreased, mineralization related genes (ENPP1 and ANKH) and calcified nodules were increased, and endoplasmic reticulum stress (ERS) was enhanced. However, ERS inhibitors could significantly inhibit the increase in calcification and the decrease in COL1A1 induced by dexamethasone. In vivo, Wistar rats received the infra-articular injection with dexamethasone (0.5 mg/kg) for 8 weeks. We found that dexamethasone treatment decreased the COL1A1 content and increased the COL2A1 content in the ACL tissues of rats and that chondroid differentiation and mineralization occurred. Meanwhile, the expression of ERS-related proteins was increased. CONCLUSION: Dexamethasone increased the calcification of ACL cells and caused ACL degeneration through ERS, suggesting that long-term treatment with dexamethasone may cause adverse effects on ACL tissue and increase the risk of long-term rupture.

GR/HDAC2/TGFßR1 pathway contributes to prenatal caffeine induced-osteoarthritis susceptibility in male adult offspring rats.

Prenatal caffeine exposure (PCE) induces developmental toxicity of multi-organ and susceptibility to multi-disease in offspring. However, the effects of PCE on osteoarthritis susceptibility in adult offspring and its intrauterine programming mechanism remain to be further investigated. Here, we found that PCE induced susceptibility to osteoarthritis in male adult offspring rats, which was related to the inhibited function of cartilage matrix synthesis from fetuses to adults. Meanwhile, PCE consistently downregulated the H3K9ac and expression levels of transforming growth factor ß receptor 1 (TGFßR1), and then blocked TGFß signaling pathway, which contributed to the suppressed cartilage matrix synthesis. Moreover, the high level of corticosterone caused by PCE reduced the H3K9ac level on TGFßR1 promoter region through acting on glucocorticoids receptor (GR) and recruiting histone deacetylase 2 (HDAC2) into the nucleus of fetal chondrocytes. Taken together, PCE induced osteoarthritis susceptibility in male adult offspring rats, which was attributed to the low-functional programming of TGFßR1 induced by corticosterone via GR/HDAC2 signaling.

Prenatal nicotine exposure increases osteoarthritis susceptibility in male elderly offspring rats via low-function programming of the TGFß signaling pathway.

Epidemiological investigations indicate that effects related to prenatal adverse environments on the organs of the offspring could continue to adulthood. This study intends to confirm that prenatal nicotine exposure (PNE) increases the susceptibility of osteoarthritis (OA) in the male offspring, and to explore the potential intrauterine programming mechanism. During pregnancy, rats were divided into a PNE group and a control group. After birth, rats were given a high-fat diet for 6 months and long-distance running for 6 weeks. The rats were euthanized at 18 months after birth (PM18) and on gestational day 20 (GD20), respectively. Knee joints were collected for histochemistry, immunohistochemistry, and quantitative polymerase chain reaction (qPCR) assays. Histological analyses and the Mankin's score showed increased cartilage destruction and accelerated OA progression in adult offspring from the PNE group. Immunohistochemistry results showed decreased expression of transforming growth factor beta (TGFß) signaling pathway. Furthermore, the expression of apoptosis factors (caspase-3 and caspase-8), inflammatory factors [interleukin (IL)-1, IL-6] and matrix degradation enzymes [matrix metalloproteinase (MMP)-3, MMP-13] were also significantly increased. Traced back to the intrauterine period, it was found that the number of chondrocytes and the contents of Col2A1 and aggrecan in the matrix in the PNE group were decreased. And, the expression of the TGFß signaling pathway was inhibited. These results suggested that PNE enhanced the susceptibility of OA in male elderly offspring rats by down-regulating TGFß signaling, which increased articular cartilage local inflammation, matrix degradation, and cell apoptosis. This study confirmed the developmental origin of OA, and clarified the congenital and the living environment impact on the occurrence and development of OA. Our findings provide a theoretical and experimental basis for OA early prevention.

Prenatal caffeine exprosure increases adult female offspring rat's susceptibility to osteoarthritis via low-functional programming of cartilage IGF-1 with histone acetylation.

Our previous in vivo studies showed that prenatal caffeine exposure (PCE) could restrain the development of chondrogenesis, which may delay fetal articular cartilage development and increase susceptibility to osteoarthritis in adults. So, the goal of the current study is to clarify theincreasing susceptibility to adult osteoarthritis in caffeine-exposed female offspring and its'mechanism. Pregnant rats were treated with 120 mg/kg·d caffeine or equal volumes of saline from gestational day (GD) 9 to 20. knee joints were collected from GD20 female fetuses and 18-week old female offspring which was treated with strenuous running for 6 weeks (55 min/day at 20 m/min) load to induce osteoarthritis. Knee joints from GD20 fetuses and adult offspring were collected for histochemistry and immunohistochemistry. Next, chondrocytes were isolated from 1-day-old newborn rats and in vitro studies were conducted where the cells in primary culture were exposed to 1, 10, and 100 µM caffeine and 250, 500, and 1,250 nM corticosterone. Insulin-like growth factor 1 (IGF-1) signal pathway genes' expression levels in fetal chondrocytes were studied, and IGF-1 histone acetylation was detected in vitro. Immunohistochemical results showed low expression levels of IGF-1 signaling genes (IGF-1, IRS-1, AKT, and COL2A1) both in fetal and adult cartilage with PCE. For adult offspring, histological results and Mankin score revealed increased cartilage destruction and accelerated osteoarthritis progression in PCE group with strenuous running exercise. Analysis in vitro revealed that caffeine and corticosterone impeded the expression of IGF-1 signaling pathway aggrecan and COL2A1 genes, but only corticosterone decreased H3K9 and H3K27 acetylation in the IGF-1 promoter region. In concluson, PCE low functional programmed cartilage IGF-1 by histone acetylation modification via overexposure to corticosterone and delayed articular cartilage development from fetus to adults. Then, the delayed cartilage development increased susceptibility to osteoarthritis in offsprings.

Role of TGFß Signaling in Maternal Ethanol-Induced Fetal Articular Cartilage Dysplasia and Adult Onset of Osteoarthritis in Male Rats.

Based on our previous findings that prenatal ethanol exposure in offspring increased susceptibility to adult osteoarthritis, this study aimed to further investigate the direct toxicity of ethanol on fetal articular cartilage development. Rat bone marrow-derived stroma cells were capsulated in alginate beads, incubated in a chondrogenic differentiation medium, and cultured for 4 weeks with ethanol treatment at concentrations of 0, 4, 20, and 100 mM. Pregnant rats were treated with ethanol (4 g/kg/day) from gestational days (GDs) 9 to 20. At GD20 and postnatal weeks 2, 6, and 12, 8 male offspring were sacrificed, and 8 male offspring rats of 8-weeks old in each group were treated with or without intraarticular injection of papain for 4 weeks to verify the susceptibility of adult osteoarthritis. Ethanol treatment resulted in poor differentiation of bone marrow-derived stroma cells to chondrocytes and suppressed the expression of the transforming growth factor-ß (TGFß)-smad2/3-Sox9 signaling pathway. In animal experiments, the shape of articular cartilage in the ethanol treatment group was more disordered than that of the control group, the matrix was not deep, and the cartilage was thin, which showed poor cartilage development. The TGFß signaling pathway in the ethanol treatment group was persistently low at all time points. After intraarticular injection of papain, histological analyses, and the Mankin score revealed increased cartilage destruction in the ethanol treatment group. Ethanol caused articular cartilage dysplasia that was programmed in adulthood via a low-functional TGFß signaling pathway, and the tolerance of this articular cartilage to external stimuli was significantly decreased.

Prenatal nicotine exposure intergenerationally programs imperfect articular cartilage via histone deacetylation through maternal lineage.

Accumulating evidence has shown that the impact of prenatal environmental factors on the organs of the offspring could last until the adulthood. Here, we aimed to investigate these effects and the potential mechanism of prenatal nicotine exposure (PNE) on the female adult cartilage of the first generation (PNE-F1) and the second generation (PNE-F2). Pregnant Wistar rats were injected with 2.0 mg/kg.d nicotine from gestational day (GD) 9 to 20. Then their F1 generation at GD20 and postnatal week (PW) 12, and F2 generation at PW12 were harvested. The expression of extracellular matrix (ECM) and transforming growth factor ß (TGFß) signaling genes were analyzed by real-time quantitative PCR, and the histone acetylation was examined by chromatin immunoprecipitation assay. The results showed that PNE reduced the ECM and TGFß signaling gene expressions in both PNE-F1 and PNE-F2 female adult articular cartilage. In the F1 generation, PNE inhibited the acetylation at H3K9 of TGFß, TGFß receptor 1 (TGFßR1), SRY-type high mobility group box 9 (SOX9), a1 chain of type II collagen (COL2A1) and aggrecan (ACAN) gene promoters at both GD20 and PW12. In PNE-F2 at PW12, the obvious deacetylation at H3K9 of the TGFßR1 and COL2A1 promoters still existed. Moreover, in rat fetal chondrocytes, corticosterone rather than nicotine directly induced the hypoacetylation of H3K9 of TGFßR1 and COL2A1 genes, which might be the main cause of imperfect cartilage for PNE-F2. This study may be helpful to elucidate the developmental variability of articular cartilage quality and useful for the early prevention of articular damage.

Early cellular responses of BMSCs genetically modified with bFGF/BMP2 co-cultured with ligament fibroblasts in a three-dimensional model in vitro.

Currently, a number of strategies including the implantation of bone marrow-derived mesenchymal stem cells (BMSCs) and growth factors have been developed to regenerate the tendon-to-bone interface after performing anterior cruciate ligament reconstruction. However, the mechanisms behind the interactions of the implanted BMSCs and tendon cells remain to be elucidated. The aim of this study was to evaluate the early cellular responses of BMSCs genetically modified with basic growth factor growth factor (bFGF)/bone morphogenic protein 2 (BMP2) and ligament fibroblasts in a three-dimensional co-culture model. BMSCs and ligament fibroblasts were both isolated from male Wistar rats. The BMSCs were then transfected with an adenoviral vector carrying bFGF or BMP2. The transfected BMSCs and ligament fibroblasts both encapsulated in alginate beads were co-cultured for 6 days in three-dimensional model. On days 0, 3 and 6, cell proliferation was assayed. On day 6, the expression of several tendon-bone related markers was evaluated. In the co-culture system, bFGF and BMP2 were highly expressed at the mRNA and protein level. During the process, bFGF significantly promoted cell proliferation, as well as the expression of scleraxis (SCX) and collagen (COL) type â…  (COL1) in the BMSCs; however, it markedly decreased the expression of phenotype markers in the ligament fibroblasts, including COL1 and COL3. BMP2 markedly increased the expression of alkaline phosphatase and osteocalcin in the BMSCs and ligament fibroblasts, whereas it had no obvious effect on cell proliferation and collagen synthesis in the ligament fibroblasts. The combination of bFGF and BMP2 resulted in the similarly enhanced proliferation of BMSCs and ligament fibroblasts as observed with bFGF alone; however, this combination more potently promoted osteogenic differentiation than did BMP2 alone. The findings of our study demonstrate the superiority of the combined use of growth factors in inducing osteogenic differentiation and provide a theoretical foundation for the regeneration of the tendon-to-bone interface.