Is Use of BMP-2 Associated with Tumor Growth and Osteoblastic Differentiation in Murine Models of Osteosarcoma?

BackgroundThe putative benefit of rhBMP-2 is in the setting of limb reconstruction using structural allografts, whether it be allograft-prosthetic composites, osteoarticular allografts, or intercalary segmental grafts. There are also potential advantages in augmenting osseointegration of uncemented endoprosthetics and in reducing infection. Recombinant human BMP-2 might mitigate nonunion in structural allograft augmented osteosarcoma limb salvage surgery; however, its use is limited because of concerns about the prooncogenic effects of the agent.Questions/purposes(1) To assess if BMP-2 signaling influences osteosarcoma cell line growth. (2) To characterize degree of osteosarcoma cell line osteoblastic differentiation in response to BMP-2. (3) To assess if BMP-2 signaling has a consistent effect on local or systemic tumor burden in various orthotopic murine models of osteosarcoma.MethodsIn this study, 143b, SaOS-2 and DLM8-M1 osteosarcoma cell lines were transfected with BMP-2 cDNA controlled by a constitutive promoter (experimental) or an empty vector (control) using a PiggyBac transposon system. Cellular proliferation was assessed using a quantitative MTT colorimetric assay. Osteoblastic differentiation was compared between control and experimental cell lines using quantitative real-time polymerase chain reaction of the osteoblastic markers connective tissue growth factor, Runx-2, Osterix, alkaline phosphatase and osteocalcin. Experimental and control cell lines were injected into the proximal tibia of either NOD-SCID (143b and SaOS-2 xenograft model), or C3H (DLM8-M1 syngeneic model) mice. Local tumor burden was quantitatively assessed using tumor volume caliper measurements and bioluminescence, and qualitatively assessed using post-mortem ex vivo microCT. Lung metastasis was qualitatively assessed by the presence of bioluminescence, and incidence was confirmed using histology. rhBMP-2 soaked absorbable collagen sponges (experimental) and sterile-H₂O soaked absorbable collagen sponges (control) were implanted adjacent to 143b proximal tibial cell line injections to compare the effects of exogenous BMP-2 application with endogenous upregulation.ResultsConstitutive expression of BMP-2 increased the in vitro proliferation of 143b cells (absorbance values 1.2 ± 0.1 versus 0.89 ± 0.1, mean difference 0.36 [95% CI 0.12 to 0.6]; p = 0.01), but had no effect on SaOS-2 and DLM8-M1 cell proliferation. In response to constitutive BMP-2 expression, 143b cells had no differences in osteoblastic differentiation, while DLM8-M1 cells downregulated the early marker connective tissue growth factor (mean ΔCt 0.2 ± 0.1 versus 0.6 ± 0.1; p = 0.002) and upregulated the early-mid range marker Runx-2 (mean ΔCt -0.8 ± 0.1 versus -1.1 ± 0.1; p = 0.002), and SaOS-2 cells upregulated the mid-range marker Osterix (mean ΔCt -2.1 ± 0.6 versus -3.9 ± 0.6; p = 0.002). Constitutive expression of BMP-2 resulted in greater 143b and DLM8-M1 local tumor volume (143b: 307.2 ± 106.8 mm³ versus 1316 ± 387.4 mm³, mean difference 1009 mm³ [95% CI 674.5 to 1343]; p < 0.001, DLM8-M1 week four: 0 mm³ versus 326.1 ± 72.8 mm³, mean difference 326.1 mm³ [95% CI 121.2 to 531]; p = 0.009), but modestly reduced local tumor growth in SaOS-2 (9.5 x 10⁸ ± 8.3x10⁸ photons/s versus 9.3 x 10⁷ ± 1.5 x 10⁸ photons/s, mean difference 8.6 x 10⁸ photons/s [95% CI 5.1 x 10⁸ to 1.2 x 10⁹]; p < 0.001). Application of exogenous rhBMP-2 also increased 143b local tumor volume (495 ± 91.9 mm³ versus 1335 ± 102.7 mm³, mean difference 840.3 mm³ [95% CI 671.7 to 1009]; p < 0.001). Incidence of lung metastases was not different between experimental or control groups for all experimental conditions.ConclusionsAs demonstrated by others, ectopic BMP-2 signaling has unpredictable effects on local tumor proliferation in murine models of osteosarcoma and does not consistently result in osteosarcoma cell line differentiation. Further investigations into other methods of safe bone and soft tissue healing augmentation and the use of differentiation therapies is warranted.Clinical RelevanceOur results indicate that BMP-2 has the potential to stimulate the growth of osteosarcoma cells that are poorly responsive to BMP-2 mediated osteoblastic differentiation. As this differentiation potential is unpredictable in the clinical setting, BMP-2 may promote the growth of microscopic residual tumor burden after resection. Our study provides further support for the recommendation to avoid the use of BMP-2 after limb-salvage surgery in patients with osteosarcoma.     

Gallbladder Polyps: What are the factors affecting the possibility of malignancy? A Single Center Experience

Hellenic Journal of Surgery - With the increasing use of imaging methods, there has been an increase in the number of patients diagnosed with gallbladder polyp. In patients undergoing...     

Intraoperative corneal thickness measurements during corneal collagen cross-linking with isotonic riboflavin solution without dextran in corneal ectasia

Objective: To monitor the changes in corneal thickness during the corneal collagen cross-linking procedure by using isotonic riboflavin solution without dextran in ectatic corneal diseases.

Materials and Methods: The corneal thickness measurements were obtained before epithelial removal, after epithelial removal, following the instillation of isotonic riboflavin solution without dextran for 30?min, and after 10?min of ultraviolet A irradiation.

Results: Eleven eyes of eleven patients with progressive keratoconus (n?=?10) and iatrogenic corneal ectasia (n?=?1) were included in this study. The mean thinnest pachymetric measurements were 391.82?±?30.34?µm (320–434?µm) after de-epithelialization of the cornea, 435?±?21.17?µm (402–472?µm) following 30?min instillation of isotonic riboflavin solution without dextran and 431.73?±?20.64?µm (387–461?µm) following 10?min of ultraviolet A irradiation to the cornea.

Conclusion: Performing corneal cross-linking procedure with isotonic riboflavin solution without dextran might not induce corneal thinning but a little swelling throughout the procedure.     

Correlation between groin pain and cup design of hip-resurfacing implants: a prospective study

Purpose

Cup design has been incriminated as the source of groin pain after hip resurfacing but has not been well described; thus, it was assessed in a prospective study looking at three implant types.

Methods

A group-match was done between three groups of hip resurfacing devices according to age, sex, body mass index, activity level, osteoarthritis aetiology and pre-operative scores.

Results

The global groin pain rate was 5.7 % at six months and 2.7 % at last follow-up. Groin pain rate was significantly different between the three groups (p = 0.004) and had a strong influence on the subjective results (p = 0.04). No groin pain emerged between six months and last follow-up. No clinical differences were noted in Harris hip score and Merle d’Aubigné-Postel score at last follow-up. However, the Oxford hip score and Devane activity score were significantly lower for cups with macrostructures.

Conclusion

The low groin pain rate in this prospective cohort was probably secondary to the specific surgical technique used and seems to be correlated with cup design. Macrostructures on the external part of the cup could be significantly harmful.     

Sensitivity and Specificity of Red Cell Distribution Width in Diagnosing Acute Mesenteric Ischemia in Patients with Abdominal Pain

Background

We investigated the utility of the red cell distribution width (RDW) in diagnosing acute mesenteric ischemia (AMI) in patients with abdominal pain.

Methods

The patients were divided into two groups in this retrospective case–control study: patients with AMI and patients with abdominal pain who did not require urgent surgery. Venous blood was collected from the patients upon admission to the emergency department, and abdominal computed tomography angiography was performed. The RDW and hematological and biochemical parameters of the groups were compared. The primary outcome was AMI among the patients with abdominal pain. The secondary outcome was mortality, complaint period, and size of ischemia/necrosis among the AMI patients.

Results

The RDW, white blood cell lactate dehydrogenase, and blood urea nitrogen of the patients with AMI were significantly different from those of the control group. When the average RDW (15.04 %) of the patients with AMI was used as a cut-off value, the sensitivity, specificity, positive likelihood ratio (+LR), and negative likelihood (?LR) were 40.8 %, 81.2 %, 2.17, and 0.73, respectively. When patients with AMI and anemia were included in the group, the sensitivity and specificity values did not change. There was no relation between the RDW and mortality, size of the ischemia/necrosis, and complaint period. Furthermore, there was no significant difference in the average RDW between the patients with ischemia/necrosis in the small intestine only and those with ischemia/necrosis in the colon.

Conclusion

The RDW on admission is of marginal help to diagnose AMI among patients with abdominal pain.     

QT dispersion predicts mortality and correlates with both coronary artery calcification and atherosclerosis in hemodialysis patients

Purpose

QT dispersion (QTd) was shown to be an independent predictor of mortality in hemodialysis (HD) patients. It may be hypothesized that coronary artery calcification is related to QTd in HD patients because widespread calcification may also involve the cardiac conducting system in these patients. In this study, we aimed to investigate the relationships of corrected QTd (QTcd) with coronary artery calcification score (CACS), carotid plaque score (CPS) and possible influence of these parameters on survival of HD patients.

Methods

Seventy-two HD patients (33 male, 39 female) were enrolled into the study. Mean age of the patients was 44 ± 12 years. Mean follow-up duration was 77 ± 24 months. CACS was determined by computed tomography. QTcd values were calculated as the difference of maximum and minimum QT intervals. Left ventricular mass index (LVMI) and CPS were measured by echocardiography.

Results

QTcd was significantly correlated with CACS (r = 0.233, p = 0.049), CPS (r = 0.354, p = 0.003) and LVMI (p = 0.011, r = 0.299). CPS was found to be significantly higher in the group with high QTcd (>60 ms) [2 (1–4) versus 0 (0–1), p = 0.02]. CACS was significantly correlated with age (r = 0.44, p < 0.001), LVMI (r = 0.52, p < 0.001) and CPS (r = 0.32, p = 0.003). In Kaplan–Meier analysis, survival of patients with high QTcd was significantly lower than the patients with low QTcd. In Cox regression analysis for predicting mortality, age, serum albumin and QTcd were found to be the independent predictors of mortality.

Conclusions

QTcd independently predicted mortality, and it was significantly associated with coronary artery calcification, left ventricular hypertrophy and atherosclerosis in HD patients.     

Comparison of intrarenal pelvic pressure during micro-percutaneous nephrolithotomy and conventional percutaneous nephrolithotomy

The micro-percutaneous nephrolithotomy (microperc) is a recently introduced percutaneous nephrolithotomy (PNL) technique that is performed through a 4.8Fr all-seeing needle. We aimed to measure the intrarenal pelvic pressure (IPP) during microperc and compare it with the levels of conventional PNL. A total of 20 patients with 1- to 3-cm renal calculi resistant to shock wave lithotripsy were treated either with microperc (Group-1, n: 10) or conventional PNL (Group-2, n: 10) by the same surgical team. The IPP was measured during different stages (entrance into the collecting system, stone fragmentation, and before termination) of the procedures by an urodynamic machine using the 6Fr ureteral catheter. All the variables were statistically compared between the two groups. The demographic values of the patients were similar. The operation time and duration of hospitalization were significantly prolonged in conventional PNL group (p = 0.034, p = 0.01, respectively). The mean drop in hematocrit levels was significantly lower in microperc group (3.5 ± 1.5 vs. 1.8 ± 0.8; p = 0.004). The IPP was significantly higher in microperc group during all steps of the procedure. The highest level of the IPP was measured as 30.3 ± 3.9 and 20.1 ± 3.1 mmHg in Group 1 and Group 2, respectively (p < 0.0001). However, the complication and success rates were found comparable. In conclusion, we demonstrate that the level of IPP is significantly increased during microperc compared to conventional PNL. Microperc should be used cautiously in cases with impaired drainage of the collecting system.