Clinicopathologic analysis of HER-2/neu immunoexpression among various histologic subtypes and grades of osteosarcoma.

Overexpression of the HER-2/neu oncogene appears to have prognostic significance in breast cancer. Recently, some have reported a relationship between increased immunohistochemical expression in osteosarcoma and poor clinical outcome. Despite limited data, a pilot trial of Herceptin, which targets the oncogene product, has been initiated for the therapy of some metastatic osteosarcomas (CCG-P9852). Archival formalin-fixed, paraffin-embedded tissue obtained from 41 patients diagnosed with osteosarcoma was examined immunohistochemically by 2 antibodies against the HER-2/neu oncogene product: CB-11 (monoclonal, 1/100) and Oncor (polyclonal, 1/200). All but one tumor (case of recurrent dedifferentiated parosteal osteosarcoma) represented primary tumor samples; when applicable, only prechemotherapy biopsies were analyzed. The study sample included the full spectrum of histologic subtypes and grades of osteosarcoma (25 conventional high grade; 3 telangiectatic; 1 small cell; 6 parosteal; 1 periosteal; and 5 low-grade intramedullary). A case of metastatic breast cancer with known overexpression of the HER-2/neu oncogene served as the positive control. Complete membranous positivity, considered prognostically significant in breast cancer, was not seen in any of our osteosarcoma cases. At least focal cytoplasmic positivity was documented in 40 (98%) tumors using the CB11 antibody and in 34 (83%) using the Oncor antibody. The intensity of the cytoplasmic staining (0, 1-3+) did not correlate with histologic subtype/grade, response to chemotherapy (<90% versus > or = 90% necrosis), metastasis, or survival. Immunohistochemical overexpression of the HER-2/neu oncogene, defined as complete membranous positivity, is not present in our series of osteosarcomas. Cytoplasmic positivity is observed in most osteosarcomas, irrespective of histologic subtype/grade, and is not associated with response to preoperative chemotherapy or disease progression.

Sex steroid receptors in Hodgkin's disease.

A case report of a dramatic therapeutic response of Hodgkin's disease (HD) to diethylstilbestrol (DES) in a man who was being treated for concurrent prostate cancer suggested that there also may be a role for sex steroids in the pathogenesis of HD (1). High levels of estrogen receptors (ER) comparable to those seen in breast carcinoma cells were detected in that patient's Hodgkin's biopsy specimen. In order to determine whether this patient was unique or whether sex steroid receptors commonly are present in HD specimens, we examined expression of ER and progesterone receptors (PR) in diagnostic tissue from pediatric (n = 14) and adult (n = 41) patients with HD using immunohistochemistry. None of the 55 samples expressed PR. 16/55 (29%) demonstrated weak nuclear ER positivity, which was confined to germinal center and occasional mantle zone lymphocytes and was comparable to that seen in non-malignant control lymph nodes. (4/5)5 (7.3%) samples exhibited moderate positivity in Reed Sternberg cells, which in one case was nuclear. ER commonly are expressed weakly in some HD tumors unrelated to clinical stage or patient sex but are generally limited to germinal center and mantle zone lymphocytes. A rare patient displays moderate cytoplasmic or nuclear ER in Reed-Sternberg cells.

Effect of intra-arterial environment on endothelialization and basement membrane organization in polytetrafluoroethylene grafts.

PURPOSE: To determine if a complete intra-arterial environment affects endothelialization rate and basement membrane organization in polytetrafluoroethylene (PTFE) grafts. METHOD: Thirty dogs underwent either infrarenal abdominal aorta PTFE interposition (12) or intraluminal stented (18) grafting. Grafts were explanted at 4 and 8 weeks and rate of endothelial ingrowth calculated. Endothelial cells were identified and basement membrane organization assessed using antibodies against endothelial cell-specific surface antigen CD31, type IV collagen, and laminin. RESULTS: Endothelialization rates, expressed as percent graft surface area coverage per week, were 3.7% +/- 0.62% (4-week control), 12.9% +/- 0.58% (4-week stented), 4.2% +/- 0.62% (8-week control), and 10.0% +/- 0.54% per week (8-week stented grafts). Endothelial repaving rates were constant for control and increased in all stented grafts (P <0.01). At 4 weeks, laminin was identified in all control (6 of 6) and no stented grafts. Staining was confined to the 20% of the hyperplastic intimal area immediately below the endothelium. At 4 weeks, type IV collagen was present throughout the entire hyperplastic intima in control specimens but was confined to a discrete subendothelial zone in stented grafts. By 8 weeks, type IV collagen became concentrated in the luminal one third of the intima in control grafts. CONCLUSION: Intra-arterial graft location is associated with early formation of an organized basement membrane and accelerated endothelialization in PTFE grafts.

Effect of balloon-expandable and self-expanding stent fixation on endoluminal polytetrafluoroethylene graft healing.

PURPOSE: To investigate the effect of stent design and deployment mechanism on endoluminal graft healing. METHOD: Twenty dogs underwent infrarenal abdominal aorta polytetrafluoroethylene (PTFE) interposition (6) or intraluminal stented grafting using either a balloon expandable (BE, n = 8) or self-expanding (SE, n = 6) stent design. Grafts were removed at 8 weeks. Length of endothelial ingrowth and intima to media height ratios (IMHR) were calculated. Perianastomotic smooth muscle (Actin+), macrophage (CD44+), proliferating (PCNA+), and platelet-derived growth factor (PDGF+) cell content were determined. RESULTS: Mean endothelial ingrowth was 1.10 +/- 0.15 cm (control), 1.88 +/- 0.13 cm (BESG), and 2.16 +/- 0.18 cm (SESG) proximally; and 0.94 +/- 0.12, 2.11 +/- 0.11 cm, and 2.16 +/- 0.15 cm, respectively, at the distal anastomosis. Endothelial ingrowth was greater in all stented grafts (P <0.001). Mean IMHRs were 1.42 +/- 0.16 (control), 0.50 +/- 0.14 (BESG), and 0.77 +/- 0.2 (SESG) proximally; and 0.84 +/- 0.1, 0.42 +/- 0.09, and 0.77 +/- 0.12 (SESG) distally. Lower IMHRs were observed in all stented graft regions (P <0.05) except the distal anastomosis of SESG. The PDGF+ and PCNA+ cell content was decreased, and Actin+ cell content was increased in all stented grafts (P <0.05). CONCLUSION: Intraluminal location enhances endothelialization and attenuates intimal thickening in PTFE grafts. The enhanced healing of intraluminal stented grafts is irrespective of the type of stent or deployment mechanism used.

The role of platelet-derived growth factor in intraluminal stented graft healing.

BACKGROUND: Intraluminally placed polytetrafluoroethylene grafts are associated with enhanced graft endothelialization and diminished intimal hyperplasia when compared with interposition grafts. This study determined the role of platelet-derived growth factor in intraluminal graft healing. STUDY DESIGN: Thirty dogs underwent infrarenal abdominal aorta polytetrafluoroethylene interposition (control, n = 15) or intraluminal stented (n = 15) grafting. Grafts were explanted at 1, 3, and 6 weeks. The percent of graft area endothelialization and intima to media height ratios were calculated. By using protein electrophoresis and the Western blot technique, platelet-derived growth factor, identified by immunolabeling with anti-platelet-derived growth factor antibody, was isolated from proximal, mid-, and distal graft regions and was quantified using densitometry. RESULTS: Graft area endothelialization was 0 +/- 3.3 percent, 2.3 +/- 3.3 percent, and 19.0 +/- 3.3 percent for 1-, 3-, and 6-week controls; and 4.7 +/- 3.7 percent, 30.5 +/- 3.3 percent, and 86.8 +/- 3.3 percent for 1-, 3-, and 6-week stented grafts. Endothelialization was greater in stented grafts at 3 and 6 weeks (p < .01). Proximal anastomosis intima to media height ratios were 1.61 +/- 0.15, 1.54 +/- 0.14, and 1.48 +/- 0.15 for 1-, 3-, and 6-week control grafts, and 0.42 +/- 0.18, 0.41 +/- 0.15, and 0.47 +/- 0.14 for 1-, 3-, and 6-week stented grafts. Similar intima to media height ratio values were present at the distal anastomosis. Lower intima to media height ratios were observed in all stented grafts (p < .01). The platelet-derived growth factor content at 1-, 3-, and 6-weeks was lower in all stented grafts when compared with controls. The content of platelet-derived growth factor was greatest in 3-week controls, with a significant difference noted in the mid-graft region (p < .05). The content of platelet-derived growth factor remained stable in all stented graft regions over 6 weeks. An inverse correlation between stented graft platelet-derived growth factor content and endothelialization (r = -0.43) and a positive correlation with proximal anastomotic intimal hyperplasia (r = 0.73) were identified. CONCLUSIONS: Lower platelet-derived growth factor content is associated with decreased intimal hyperplasia and improved healing in intra-arterial polytetrafluoroethylene grafts.

Effect of endoluminal PTFE graft placement on cell proliferation, PDGF secretion, and intimal hyperplasia.

To determine the effects of intraluminal placement on peri-anastomotic intimal hyperplasia and platelet derived growth factor (PDGF) secretion in polytetrafluoroethylene (PTFE) grafts. Infrarenal aortic PTFE grafts were placed in 30 dogs as either interposition (n = 12) or intraluminal stented (n = 18) grafts. Grafts were explanted at 4 and 8 weeks. At each anastomosis, intima to media height ratios (IMHR) were calculated, and smooth muscle (Actin), proliferating (PCNA), and PDGF secreting cell content determined using cell-specific immunohistochemical stains. At the proximal anastomosis, control and stented graft IMHRs were 1.01 +/- 0.16 vs 0.59 +/- 0.18 in 4-week and 1.42 +/- 0.16 vs 0.50 +/- 0.14 in 8-week specimens. Similar IMHR values were present for the distal anastomosis. Peri-anastomotic PCNA cell counts were greater in control grafts at both 4 and 8 weeks. Stented grafts were associated with diminished IMHR and PCNA+ content at both 4 and 8 weeks (P < 0.05). PDGF+ content was similar among control and stented grafts at 4 weeks, while lower in stented grafts at 8 weeks (P < 0.05). At the distal anastomosis, a correlation between PDGF secretion and Actin+ cell content was observed in control grafts at 4 (r = 0.74) and 8 (r = -0.56) weeks. Cell proliferation was associated with PDGF content in 4-week intraluminal and 8-week control grafts (P < 0.05). Changes in IMHR were not the result of differential PDGF secretion. Intraluminal location attenuates intimal hyperplasia in PTFE grafts. The reduced intimal hyperplasia and improved healing of endoluminal grafts could not be attributed to lower PDGF content alone.