Rapamycin increases pCREB, Bcl-2, and VEGF-A through ERK under normoxia.

Rapamycin may serve as a new anti-osteosarcoma (OSA) agent due to its ability to inhibit the metastatic behavior of OSA. However, only limited benefit is observed in rodent studies and clinical trials using rapamycin as a single agent in the treatment of OSA. The target of rapamycin, mammalian target of rapamycin has multiple biological functions and may be linked with the kinases that mediate the phosphorylation of cyclic AMP-responsive element-binding (CREB) protein, an import factor in tumor progression. By employing an OSA cell line MG-63, we investigated how rapamycin regulates the phosphorylation of CREB (pCREB) at Ser133 and the expressions of two putative CREB targets, B-cell lymphoma 2 (Bcl-2) and vascular endothelial growth factor-A (VEGF-A). Under normoxia, we found that rapamycin (100 nM) induced an increase of pCREB that was prevented by mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) inhibitor U0126 or cAMP-dependent protein kinase (PKA) inhibitor H89. However, H89 enhanced Akt phosphorylation and did not decrease the cell viability upon rapamycin treatment. In contrast, U0126 did not enhance Akt phosphorylation and decreased the cell viability upon rapamycin treatment. Moreover, U0126 prevented the rapamycin-induced increase of Bcl-2 and VEGF-A levels. Under hypoxia, rapamycin effectively prevented the hypoxia-induced increase of pCREB, Bcl-2, and VEGF-A. Our study demonstrated that rapamycin might be less effective in treating OSA cells under normoxia and provided the rationale for a combination of rapamycin and MEK/ERK inhibitor in the treatment of OSA.

Anterior surgical options for the treatment of cervical spondylotic myelopathy in a long-term follow-up study.

OBJECTIVE: To provide a basis for the choice of anterior surgery procedures in the treatment of cervical spondylotic myelopathy (CSM) through long-term follow-up. METHODS: A consecutive series of 89 patients with CSM having complete follow-up data were analyzed retrospectively. All patients were treated with anterior cervical discectomy and fusion (ACDF), and anterior cervical corpectomy and fusion (ACCF) from July 2000 to June 2007. The lesions were located in one segment (n = 25), two segments (n = 56), and three segments (n = 8). Preoperative and postoperative, the C2-C7 angle, cervical intervertebral height, radiographic fusion status, result of the adjacent segment degeneration, the Japanese Orthopaedic Association (JOA), and the Short Form 36-item (SF36) questionnaire scores were used to evaluate the efficacy of the surgery. RESULTS: According to the different compression conditions of the 89 cases, different anterior operation procedures were chosen and satisfactory results were achieved, indicating that direct anterior decompressions were thorough and effective. The follow-up period was 60-108 months, and the average was 79.6 months. The 5-year average symptom improvement rate, effectiveness rate, and fineness rate were 78.36 %, 100 % (89/89), and 86.52 % (77/89), respectively. CONCLUSIONS: For CSM with compression coming from the front side, proper anterior decompression based on the specific conditions could directly eliminate the compression. Through long-term follow-up, the effect of decompression became observable.

Bone induction by biomimetic PLGA copolymer loaded with a novel synthetic RADA16-P24 peptide in vivo.

Bone morphogenetic protein-2 (BMP-2) is a key bone morphogenetic protein, and poly(lactic-co-glycolic acid) (PLGA) has been widely used as scaffold for clinical use to carry treatment protein. In the previous studies, we have synthesized BMP-2-related peptide (P24) and found its capacity of inducing bone regeneration. In this research, we have synthesized a new amphiphilic peptide Ac-RADA RADA RADA RADA S[PO4]KIPKASSVPTELSAISTLYLDDD-CONH2 (RADA16-P24) with an assembly peptide RADA16-Ion the P24 item of BMP2 to form divalent ion-induced gelatin. Two methods of physisorption and chemical cross-linking were used to bind RADA16-P24 onto the surface of the copolymer PLGA to synthesize RADA16-P24-PLGA, and its capacity of attaching bone marrow stromal cells (BMSCs) was evaluated in vitro and inducing ectopic bone formation was examined in vivo. In vitro our results demonstrated that RADA16-P24-PLGA copolymer prepared by physisorbing or prepared by chemical cross-linking had a peptide binding rate of (2.0180±0.5296)% or (10.0820±0.8405)% respectively (P<0.05). In addition the BMSCs proliferated vigorously in the RADA16-P24-PLGA biomaterials. Significantly the percentage of BMSCs attached to RADA16-P24-PLGA composite prepared by chemical cross-linking and physisorbing were (71.4±7.5) % or (46.7±5.8) % (P<0.05). The in vivo study showed that RADA16-P24-PLGA chemical cross-linking could better induce ectopic bone formation compared with RADA16-P24-PLGA physisorbing and PLGA. It is concluded that the PLGA copolymer is a good RADA16-P24 carrier. This novel RADA16-P24-PLGA composite has strong osteogenic capability.