Learning Curve and Clinical Outcomes of Ultrasonic Osteotome-based En Bloc Laminectomy for Thoracic Ossification of the Ligamentum Flavum.

OBJECTIVE: Despite rapid advances in minimally invasive surgery, en bloc laminectomy remains the most common surgical approach for treating thoracic ossification of the ligamentum flavum (TOLF). However, the learning curve of this risky operation is rarely reported. Therefore, we aimed to describe and analyze the learning curve of ultrasonic osteotome-based en bloc laminectomy for TOLF. METHODS: Among 151 consecutive patients with TOLF who underwent en bloc laminectomy performed by one surgeon between January 2012 and December 2017, we retrospectively analyzed their demographic data, surgical parameters, and neurological function. Neurological outcome was evaluated with the modified Japanese Orthopaedic Association (mJOA) scale, and the Hirabayashi method was used to calculate the neurological recovery rate. The learning curve was assessed with logarithmic curve-fitting regression analysis. Univariate analysis methods were used for statistical analysis, including t-test, rank sum test, and chi-square test. RESULTS: A total of 50% of learning milestones could be reached in approximately 14 cases, and the asymptote in 76 cases. Therefore, 76 of the 151 enrolled patients were defined as the "early group," and the remaining 75 were delimitated as the "late group" for comparison. There was a significant intergroup difference in the corrected operative time (94.80 ± 27.77 vs 65.93 ± 15.67 min, P < 0.001) and the estimated blood loss (median 240 vs 400 mL, P < 0.001). The overall follow-up was 83.1 ± 18.5 months. The mJOA significantly increased from a median of 5 (IQR: 4-5) before the surgery to 10 (IQR: 9-10) at the last follow-up (P < 0.001). The overall complication rate was 37.1%, and no significant intergroup difference was found, except for the incidence of dural tears (31.6% vs 17.3%, p = 0.042). CONCLUSION: Initially, mastering the en bloc laminectomy technique using ultrasonic osteotome for TOLF treatment can be challenging, but the surgeon's experience improves as the operative time and blood loss decrease. Improved surgical experience reduced the risk of dural tears but was not associated with the overall complication rate or long-term neurological function. Despite the relatively long learning curve, en bloc laminectomy is a secure and valid technique for TOLF treatment.

Incidence, clinical features, and risk factors for acute pancreatitis following posterior instrumented fusion surgery for lumbar degenerative disease: a single-center, retrospective analysis of 20,929 patients.

PURPOSE: The aim of this study is to identify the incidence, clinical features, and risk factors for postoperative acute pancreatitis (PAP) after lumbar surgery. METHODS: We retrospectively analyzed patients who developed PAP after posterior lumbar fusion surgery. For each PAP patient, data were collected for four controls who underwent procedures in the same period and did not develop PAP. Statistical methods included univariate and multivariate analyses. RESULTS: Totally, 21 out of 20,929 patients were diagnosed with PAP (0.10%) after posterior lumbar fusion surgery. Patients with degenerative lumbar scoliosis were at higher risk of developing PAP (P < 0.05). With atypical clinical features, PAP occurred within 3 days (0-5) after surgery. PAP patients had significantly higher incidence of osteoporosis (47.6 vs. 22.6%, P = 0.030) and fusion of L1/2(42.9 vs. 4.3%, P = 0.010), lower albumin (42.2 ± 4.1 vs. 44.3 ± 3.2 g/L, P = 0.010), more fusion segments (median 4 vs. 3, P = 0.022), larger surgical invasiveness index (median 9 vs. 8, P = 0.007), longer operation duration (232 ± 109 vs. 185 ± 90 min, P = 0.041), greater estimated blood loss (median 600 vs. 400 mL, P = 0.025), lower intraoperative mean arterial pressure (87.2 ± 9.9 vs. 92.1 ± 8.8 mmHg, P = 0.024). Multivariate logistic regression analysis found three independent risk factors: fusion of L1/2, surgical invasiveness index > 8, and intraoperative mean arterial pressure < 90 mmHg. All patients were treated with conservative therapy and fully recovered after 8.1 (4-22) days. CONCLUSION: The incidence of PAP following posterior surgery for degenerative lumbar disease was 0.10%, and its clinical features were not typical. The fusion of L1/2, high surgical invasiveness index, and low intraoperative mean arterial pressure were independent risk factors for PAP after surgery for lumbar degenerative disease.

Development and validation of a novel thoracic spinal stenosis surgical invasiveness index: a single-center study based on 989 patients.

BACKGROUND CONTEXT: Surgical invasiveness indices have been established for general spine surgery (surgical invasiveness index [SII]), spine deformity, and metastatic spine tumors; however, a specific index for thoracic spinal stenosis (TSS) has not been developed. PURPOSE: To develop and validate a novel invasiveness index, incorporating TSS-specific factors for open posterior TSS surgery, which may facilitate the prediction of operative duration and intraoperative blood loss, and the stratification of surgical risk. STUDY DESIGN: A retrospective observational study. PATIENT SAMPLE: Overall, 989 patients who underwent open posterior TSS surgeries at our institution during the past 5 years were included. OUTCOME MEASURES: The operation duration, estimated blood loss, transfusion status, major surgical complications, length of hospital stay, and medical expenses. METHODS: We retrospectively analyzed the data of 989 consecutive patients who underwent posterior surgery for TSS between March 2017 and February 2022. Among them, 70% (n=692) were randomly placed in a training cohort, and the remaining 30% (n=297) automatically constituted the validation cohort. Multivariate linear regression models of operative time and log-transformed estimated blood loss were created using TSS-specific factors. Beta coefficients derived from these models were used to construct a TSS invasiveness index (TII). The ability of the TII to predict surgical invasiveness was compared with that of the SII and assessed in a validation cohort. RESULTS: The TII was more strongly correlated with operative time and estimated blood loss (p<.05) and explained more variability in operative time and estimated blood loss than the SII (p<.05). The TII explained 64.2% of operative time and 34.6% of estimated blood loss variation, whereas the SII explained 38.7% and 22.5%, respectively. In further verification, the TII was more strongly associated with transfusion rate, drainage time, and length of hospital stay than SII (p<.05). CONCLUSIONS: By incorporating TSS-specific components, the newly developed TII more accurately predicts the invasiveness of open posterior TSS surgery than the previous index.

Thirty-day unplanned reoperations of thoracic spine surgery: 10 years of data from a single center with 3242 patients.

BACKGROUND CONTEXT: Unplanned reoperation is a useful quality indicator for spine surgery. However, the rates of a 30-day unplanned reoperation in patients undergoing thoracic spinal surgery are not well established. PURPOSE: To assess the rates, reasons, and risk factors of 30-day unplanned reoperations for thoracic spine surgeries in a single center study. STUDY DESIGN: A retrospective observational study. PATIENT SAMPLE: A total of 3242 patients who underwent thoracic spinal surgery at our institution in the past decade were included. OUTCOME MEASURES: The incidence, chief reasons, and risk factors for unplanned reoperations within 30 days after thoracic spinal surgery. METHODS: We retrospectively analyzed the data of all patients who underwent thoracic spinal surgery between January 2012 and December 2021. Statistical methods, including univariate and multivariate analyses, were performed to assess the incidence, reasons, and risk factors for thoracic degenerative diseases, spinal tumors, kyphosis deformity, and spinal trauma. RESULTS: Of the 3242 patients who underwent thoracic spinal surgery, 107 (3.30%) required unplanned reoperations within 30 days due to epidural hematoma (1.17%), wound complications (0.80%), implant complications (0.43%), inadequate decompression (0.25%), and other causes (0.65%). Patients with degenerative disease (3.88%), spinal tumor (2.98%), and kyphosis deformity (3.33%) had significantly higher incidences of reoperation than those with spinal trauma (1.47%). Unplanned reoperations were classified as hyperacute (30.84%), acute (31.76%), and subacute (37.38%). After univariate analysis, several factors were associated with unplanned reoperation in the 4 cohorts of thoracic spine diseases (p<.05). Multivariate logistic regression analysis revealed that upper thoracic spine surgery (p=.001), concomitant dekyphosis (p=.027), and longer activated partial thromboplastin time (p=.025) were risk factors of unplanned reoperation for thoracic degenerative disease. Whereas American Society of Anesthesiologists (ASA) grade III (p=.015), combined approach (p=.016), and operation time longer than 420 min (p=.042) for spinal tumor, and similar ankylosing spondylitis (p=.023) and operation time longer than 340 min (p=.041) were risk factors of unplanned reoperation for kyphosis deformity. CONCLUSIONS: The unplanned reoperation rate for thoracic spine surgery was 3.30%, with epidural hematoma and wound complications being the most common reasons. However, upper thoracic spine surgery, concomitant dekyphosis, underlying coagulation disorder, longer operation time, higher ASA grade, and comorbidities of ankylosing spondylitis led to an increased risk of unplanned reoperation within 30 days of thoracic spine surgery.

Thirty-day Unplanned Reoperations After Posterior Surgery for Thoracic Spinal Stenosis: A Single-center Study Based on 1948 Patients.

STUDY DESIGN: A retrospective study. OBJECTIVE: The purpose of this study is to identify the incidences, causes, and risk factors of 30-day unplanned reoperation of posterior surgery for thoracic spinal stenosis (TSS) based on 1948 patients in a single center. SUMMARY OF BACKGROUND DATA: Unplanned reoperation is suggested to be a useful quality indicator for spine surgery. However, the incidences, causes, and risk factors of 30-day unplanned reoperation in patients who underwent posterior spinal surgery for TSS have not been well-established. MATERIALS AND METHODS: We retrospectively analyzed the clinical data of patients who underwent posterior spinal surgery for TSS from January 2011 to December 2021. Statistical methods including univariate and multivariate analyses were performed to assess the incidences, causes, and risk factors. RESULTS: A total of 1948 patients who underwent posterior spinal surgery for TSS in our institution were reviewed, and 77 (3.95%) required unplanned reoperations within 30 days because of epidural hematoma (1.64%), wound-related complications (1.02%), inadequate decompression (0.41%), and implant malposition or failure (0.36%), neurological deficit (0.26%), and other causes (0.26%). After univariate analysis, seven clinical factors were associated with unplanned reoperation ( P <0.05). Multivariate logistic regression analysis showed that upper thoracic spine surgery ( P =0.010), thoracic kyphosis ≥45° ( P =0.039), and intraoperative dural injury ( P =0.047) were independent risk factors for 30-day unplanned reoperation of posterior surgery for TSS. CONCLUSIONS: The incidence of 30-day unplanned reoperations after posterior surgical treatment for TSS was 3.95%. The most common causes were epidural hematoma, wound-related complications, inadequate decompression, and implant malposition or failure. Upper thoracic spine surgery, thoracic kyphosis ≥45°, and intraoperative dural injury led to an increased risk of unplanned reoperation within 30 days after posterior spinal surgery for TSS. LEVEL OF EVIDENCE: 4.

Incidences, causes and risk factors of unplanned reoperations within 30 days of spine surgery: a single-center study based on 35,246 patients.

BACKGROUND CONTEXT: Unplanned reoperation, a quality indicator in spine surgery, has not been sufficiently investigated in a large-scale, single-center study. PURPOSE: To assess the incidences, causes, and risk factors of unplanned reoperations within 30 days of spine surgeries in a single-center study. STUDY DESIGN: Retrospective observational study. PATIENT SAMPLE: A cohort of 35,246 patients who underwent spinal surgery in our hospital were included. OUTCOME MEASURES: The rates, chief reasons, and risk factors for unplanned reoperations within 30 days of spine surgery. METHODS: We retrospectively analyzed the data for patients who underwent spine surgeries for degenerative spinal disorders, tumor, or deformity and had subsequent unplanned operations within 30 days at a single tertiary academic hospital from January 2016 to July 2021. Univariate and multivariate analyses were performed to assess the incidences, causes, and risk factors. RESULTS: Out of 35,246 spinal surgery patients, 297 (0.84%) required unplanned reoperations within 30 days of spine surgery. Patients with a thoracic degenerative disease (3.23%), spinal tumor (1.63%), and spinal deformity (1.50%) had significantly higher rates of reoperation than those with atlantoaxial (0.61%), cervical (0.65%), and lumbar (0.82%) degenerative disease. The common causes for reoperation included epidural hematoma (0.403%), wound infections (0.148%), neurological deficit (0.108%), and pedicle screw malposition (0.077%). Unplanned reoperations were classified as hyperacute (45.45%), acute (30.98%), subacute (15.82%), or chronic (7.74%). Univariate analysis indicated that 20 clinical factors were associated with unplanned reoperation (p<.05). Multivariate Poisson regression analysis revealed that anemia (p<.001), osteoporosis (p=.048), ankylosing spondylitis (p=.008), preoperative foot drop (p=.011), deep venous thrombosis (p<.001), and previous surgical history (p<.001) were independent risk factors for unplanned spinal reoperation. CONCLUSIONS: The incidence of unplanned spinal reoperations was 0.84%. The chief common causes were epidural hematoma, wound infections, neurological deficit, and pedicle screw malposition. Anemia, osteoporosis, ankylosing spondylitis, preoperative foot drop, deep venous thrombosis, and previous surgical history led to an increased risk of unplanned reoperation within 30 days of spine surgery.

Does Early Graft Patency Benefit from Perioperative Statin Therapy? A Propensity Score-Matched Study of Patients Undergoing Off-Pump Coronary Artery Bypass Surgery.

BACKGROUND: Decreased graft patency after off-pump coronary artery bypass grafting (OPCAB) leads to substantial increases in cardiac events. However, there is paucity of data on efficacy and safety of perioperative statin therapy for OPCAB populations. METHODS: 582 patients undergoing OPCAB in a single-institution database (October 1, 2009-September 30, 2012) were stratified by perioperative continuation of statin therapy (CS group, n=398) or not (DS group, n=184). Inverse probability weighted propensity adjustment was used to account for treatment assignment bias, resulting in a well-matched cohort. Primary outcomes were graft patency at an average of five days after operation and in-hospital mortality. Secondary outcomes included intraoperative blood loss, liver, and renal functions. RESULTS: No in-hospital death occurred in this study. Early graft patency rates after OPCAB were 98.4% (1255 of 1275 grafts) in the CS group and 98.0% (583 of 595 grafts, P=0.486) in the DS group. Secondary outcomes showed a reduction in blood loss during operation (438.53 mL versus 480.47 mL, P=0.01). Continuation of statin therapy is associated with alanine transaminase (ALT) elevation (49.67 U/L versus 34.52 U/L, P<0.001), as well as aspartate transaminase (33.54 U/L versus 28.10 U/L, P<0.001). Abnormal ALT elevation was observed in 8.9% of the CS group and 3.1% in DS (odds ratio 3.06, 95% confidence interval, 1.77 to 5.29, P<0.001). There was no significant difference in estimated glomerular filtration rate (76.28 mL/min/1.73m2 versus 76.13 mL/min/1.73m2, P=0.90). Subgroup analyses suggested that graft occlusion was less common in CS than in DS group among smoking patients (odds ratio 0.41, 95% confidence interval, 0.20 to 0.86, P=0.026). CONCLUSIONS: Perioperative continuation of statin therapy did not improve early graft patency in OPCAB patients. A lower risk of graft occlusion was observed among smoking patients. Continuous statin use correlated with liver function elevation (Clinical Trials.gov number, NCT01268917).

Mutational spectrum of Barrett's stem cells suggests paths to initiation of a precancerous lesion.

The precancerous lesion known as Barrett's oesophagus can evolve to oesophageal adenocarcinoma in decades-long processes of regenerative growth. Here we report the isolation and propagation of distinct, patient-matched stem cells of Barrett's, gastric and oesophageal epithelia that yield divergent tumour types following in vitro transformation and xenografting. Genomic analyses reveal a broad mutational spectrum unique to Barrett's stem cells that likely reflects their risk for oncogenesis. Remarkably, 25% of cases show no cancer-related genomic changes, suggesting that Barrett's initiates without driver mutations. Most cases, however, sustain patterns of deletions almost identical to adenocarcinoma though tumour-associated gene amplifications were absent. Notably, those suspected of low-grade dysplasia have p53 mutations or undergo amplifications of proto-oncogenes and receptor tyrosine kinases, implicating these events in lethal transitions. Our findings suggest paths for the initiation and progression of Barrett's and define a discrete stem cell underlying its regenerative growth whose eradication could prevent oesophageal adenocarcinoma.

c-Abl promotes osteoblast expansion by differentially regulating canonical and non-canonical BMP pathways and p16INK4a expression.

Defects in stem cell renewal or progenitor cell expansion underlie ageing-related diseases such as osteoporosis. Yet much remains unclear about the mechanisms regulating progenitor expansion. Here we show that the tyrosine kinase c-Abl plays an important role in osteoprogenitor expansion. c-Abl interacts with and phosphorylates BMPRIA and the phosphorylation differentially influences the interaction of BMPRIA with BMPRII and the Tab1-Tak1 complex, leading to uneven activation of Smad1/5/8 and Erk1/2, the canonical and non-canonical BMP pathways that direct the expression of p16(INK4a). c-Abl deficiency shunts BMP signalling from Smad1/5/8 to Erk1/2, leading to p16(INK4a) upregulation and osteoblast senescence. Mouse genetic studies revealed that p16(INK4a) controls mesenchymal stem cell maintenance and osteoblast expansion and mediates the effects of c-Abl deficiency on osteoblast expansion and bone formation. These findings identify c-Abl as a regulator of BMP signalling pathways and uncover a role for c-Abl in p16(INK4a) expression and osteoprogenitor expansion.

A crucial role for bone morphogenetic protein-Smad1 signalling in the DNA damage response.

DNA damage and the elicited cellular response underlie the etiology of tumorigenesis and ageing. Yet, how this response integrates inputs from cells' environmental cues remains underexplored. Here we report that the BMP-Smad1 pathway, which is essential for embryonic development and tissue homeostasis, has an important role in the DNA damage response and oncogenesis. On genotoxic stress, Atm phosphorylates BMPs-activated Smad1 in the nucleus on S239, which disrupts Smad1 interaction with protein phosphatase PPM1A, leading to enhanced activation and upregulation of Smad1. Smad1 then interacts with p53 and inhibits Mdm2-mediated p53 ubiquitination and degradation to regulate cell proliferation and survival. Enhanced Smad1 S239 phosphorylation, and Smad1 mutations causing S239 substitution were detected in oesophageal and gastric cancer samples, respectively. These findings suggest that BMP-Smad1 signalling participates in the DNA damage response via the Atm-p53 pathway, thus providing a molecular mechanism whereby BMP-Smad1 loss-of-function leads to tumorigenesis, for example, juvenile polyposis and Cowden syndromes.

PKCdelta mediates Nrf2-dependent protection of neuronal cells from NO-induced apoptosis.

A chemical inhibitor library of 84 compounds was screened to investigate the signaling pathway(s) leading to activation of Nrf2 in response to nitric oxide (NO). We identified the protein kinase C delta (PKCdelta) inhibitor rottlerin as the only compound that reduced NO-induced ARE-luciferase reporter activity and diminished NO-induced up-regulation of two Nrf2/ARE-regulated proteins - NAD(P)H:quinone oxidoreductase-1 (NQO1) and hemeoxygenase-1 (HO-1) in SH-Sy5y cells. Rottlerin also sensitized neuroblastoma cells and mouse primary cortical neurons to NO-induced apoptosis. Stable over-expression of PKCdelta augmented NO-induced, ARE-dependent gene expression of HO-1 in SH-Sy5y cells, which were more protected from NO killing. Conversely, NO-induced ARE-dependent gene expression was reduced in PKCdelta-knockdown SH-EP cells, which displayed greater sensitivity to apoptosis. PKCdelta(-/-) cortical neurons exhibited increased NO-induced apoptosis and less HO-1 mRNA and protein induction compared with wild type neurons. Hence, PKCdelta is an important positive modulator of NO-induced Nrf2/ARE-dependent signaling that counteracts NO-mediated apoptosis in neuronal cells.

p53 functions as a negative regulator of osteoblastogenesis, osteoblast-dependent osteoclastogenesis, and bone remodeling.

p53 is a well known tumor suppressor. We show that p53 also regulates osteoblast differentiation, bone formation, and osteoblast-dependent osteoclast differentiation. Indeed, p53(-/-) mice display a high bone mass phenotype, and p53(-/-) osteoblasts show accelerated differentiation, secondary to an increase in expression of the osteoblast differentiation factor osterix, as a result. Reporter assays indicate that p53 represses osterix transcription by the minimal promoter in a DNA-binding-independent manner. In addition, p53(-/-) osteoblasts have an enhanced ability to favor osteoclast differentiation, in association with an increase in expression of macrophage-colony stimulating factor, which is under the control of osterix. Furthermore, inactivating p53 is sufficient to rescue the osteoblast differentiation defects observed in mice lacking c-Abl, a p53-interacting protein. Thus, these results identify p53 as a novel regulator of osteoblast differentiation, osteoblast-dependent osteoclastogenesis, and bone remodeling.

Identification of TopBP1 as a c-Abl-interacting protein and a repressor for c-Abl expression.

Expression of BCR-ABL is the leading cause of chronic myelogenous leukemia. In chronic myelogenous leukemia cells, c-Abl expression is silenced by promoter methylation. In addition, the level of c-Abl needs to be tightly and constantly regulated due to its cytotoxicity and its rapid degradation after activation. Yet the regulation of c-Abl expression remains unclear. In an effort to gain better understanding of c-Abl function, we performed a glutathione S-transferase-Abl pull-down screen and identified TopBP1, a topoisomerase IIbeta-binding protein that contains Brca1 C-terminal motifs and has been implicated in DNA damage response. Their physical interaction was verified by in vitro and in vivo assays with TopBP1 found as a substrate of Abl proteins. TopBP1 could repress the expression of c-Abl at both mRNA and protein levels. Reporter assays indicate that TopBP1 directly repressed the promoter activity of c-Abl. Furthermore, TopBP1 repressed expression of c-Abl through a novel mechanism that involved histone deacetylation and DNA methylation. This transcriptional repression was inhibited by c-Abl in a kinase-dependent manner. The dual antagonistic interplay between c-Abl and TopBP1 may also provide a mechanism for fine-tuning of c-Abl levels.

ERK phosphorylates p66shcA on Ser36 and subsequently regulates p27kip1 expression via the Akt-FOXO3a pathway: implication of p27kip1 in cell response to oxidative stress.

Mice deficient for p66shcA represent an animal model to link oxidative stress and aging. p66shcA is implicated in oxidative stress response and mitogenic signaling. Phosphorylation of p66shcA on Ser36 is critical for its function in oxidative stress response. Here we report the identification of ERK as the kinase phosphorylating p66shcA on Ser36. Activation of ERKs was necessary and sufficient for Ser36 phosphorylation. p66shcA interacted with ERK and was demonstrated to be a substrate for ERK, with Ser36 being the major phosphorylation site. Furthermore, in response to H2O2, inhibition of ERK activation repressed p66shcA-dependent phosphorylation of FOXO3a and the down-regulation of its target gene p27kip1. Down-regulation of p27 might promote cell survival, as p27 played a proapoptotic role in oxidative stress response. As a feedback regulation, Ser36 phosphorylated p66shcA attenuated H2O2-induced ERK activation, whereas p52/46shcA facilitated ERK activation, which required tyrosine phosphorylation of CH1 domain. p66shcA formed a complex with p52/46ShcA, which may provide a platform for efficient signal propagation. Taken together, the data suggest there exists an interplay between ERK and ShcA proteins, which modulates the expression of p27 and cell response to oxidative stress.

Distinct roles of c-Abl and Atm in oxidative stress response are mediated by protein kinase C delta.

c-Abl and Atm have been implicated in cell responses to DNA damage and oxidative stress. However, the molecular mechanisms by which they regulate oxidative stress response remain unclear. In this report, we show that deficiency of c-Abl and deficiency of ATM differentially altered cell responses to oxidative stress by induction of antioxidant protein peroxiredoxin I (Prx I) via Nrf2 and cell death, both of which required protein kinase C (PKC) delta activation and were mediated by reactive oxygen species. c-abl-/- osteoblasts displayed enhanced Prx I induction, elevated Nrf2 levels, and hypersusceptibility to arsenate, which were reinstated by reconstitution of c-Abl; Atm-/- osteoblasts showed the opposite. These phenotypes correlated with increased PKC delta expression in c-abl-/- osteoblasts and decreased PKC delta expression in Atm-/- cells, respectively. The enhanced responses of c-abl-/- osteoblasts could be mimicked by overexpression of PKC delta in normal cells and impeded by inhibition of PKC delta, and diminished responses of Atm-/- cells could be rescued by PKC delta overexpression, indicating that PKC delta mediated the effects of c-Abl and ATM in oxidative stress response. Hence, our results unveiled a previously unrecognized mechanism by which c-Abl and Atm participate in oxidative stress response.

Activation of p38 mitogen-activated protein kinase is required for osteoblast differentiation.

p38 MAPK is a conserved subfamily of MAPKs involved in inflammatory response, stress response, cell growth and survival, as well as differentiation of a variety of cell types. In this report we demonstrated that p38 MAPK played an important role in osteoblast differentiation using primary calvarial osteoblast, bone marrow osteoprecursor culture, and a murine cell line, MC3T3-E1. We found that p38 MAPK was activated as calvarial osteoblast differentiates along with extracellular signal-regulated kinases (ERKs). When p38 MAPK is inhibited with a specific inhibitor, the expression of differentiation markers, such as alkaline phosphatase and mineral deposition, were significantly reduced. MC3T3-E1 cells expressing dominant negative p38 MAPK also displayed signs of delay in ALP and mineral deposition. Differentiation of the bone marrow osteoprecursors was also impeded by the p38 MAPK inhibitor, justified by the same markers. Yet the inhibitory effects observed in calvarial osteoblasts and bone marrow osteoprogenitor cells could be partially prevailed by bone morphogenetic protein-2. Inhibition of ERKs with a specific drug did not significantly affect osteoblast differentiation even though ERK1/2 were also activated during osteoblast differentiation. These results taken together indicate that p38 MAPK, but not ERKs, is necessary for osteoblast differentiation.

Chromatin-remodelling factor BRG1 selectively activates a subset of interferon-alpha-inducible genes.

Brahma-related gene 1 (BRG1 ) is a key component of the ATP-dependent chromatin-remodelling SWI2-SNF2 complex and has been implicated in regulating gene expression, cell-cycle control and tumorigenesis. Here we report that BRG1 interacts with signal transducer and activator of transcription 2 (STAT2) - a transcription factor that regulates gene expression mediated by interferon-alpha (IFN-alpha). BRG1 enhances the IFN-alpha-induced expression of 9-27 and IFI27 but not that of four other target genes tested, showing that the activation of different target genes by STAT2 may involve alternative chromatin modifiers. Our results also suggest that the recruitment and activation of BRG1 may require other cis-acting and trans-acting elements in addition to STAT2. Our study links the SWI2-SNF2 complex to the regulation of cytokine-induced gene expression and may identify a molecular mechanism of BRG1-mediated gene activation and tumorigenesis.