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Eugene A Woltering

Publications and source records attributed to Eugene A Woltering.

At least 19 recordsLinked to original sources
PubMed2006

Development of 125I-methylene blue for sentinel lymph node biopsy.

BACKGROUND: Sentinel lymph node biopsy is an established alternative to complete lymph node dissection in some patients. We have developed a novel, radiolabeled methylene blue dye that may be a useful alternative to the traditional two-step procedure involving 99mTc-labeled colloid and unlabeled blue dye. We hypothesize that 125I-labeled methylene blue will be rapidly absorbed into the lymphatics and transported to the drainage basin containing the sentinel nodes. MATERIALS AND METHODS: Rabbits footpads were injected with 1 mCi of 125I-labeled methylene blue admixed with unlabeled dye. A hand-held gamma detection device allowed tracking of radiolabeled dye to nodes in the popliteal and inguinal regions. At pre-established time points animals were sacrificed, and the nodal basin dissected. Nodal radioactivity as well as uptake of blue dye was recorded. RESULTS: The spread of the radiolabeled methylene blue compound from the footpad to the popliteal lymph nodes occurred in 5-10 min. CONCLUSION: The radiolabeled dye rapidly progresses through lymphatics to the draining nodes. Use of radiolabeled methylene blue may be an attractive alternative to current two-step sentinel node techniques, as it may be less painful, and may reduce the cost associated with the time-delay between the injection of the radioactive compound and surgery.

Animals↗
PubMed2006

Gallic acid is partially responsible for the antiangiogenic activities of Rubus leaf extract.

An aqueous extract of leaves from Rubus suavissimus S. Lee (Rosaceae) or sweet leaf tea was tested for antiangiogenic activity in a human tissue-based fibrin-thrombin clot angiogenesis assay. Further fractionation of this crude extract was performed and the antiangiogenic effect of individual fractions was assessed. The extract was also tested for its oral bioavailability by using the serum of normal rats gavaged with the extract in the assay. At a 0.1% w/v concentration, the extract inhibited initiation of the angiogenic response and subsequent neovessel growth from samples that had already initiated an angiogenic response. Two subfractions of the extract showed significant inhibition of angiogenesis at 0.1% w/w. Gallic acid was elucidated as one of the active angiogenesis inhibitors in one fraction. A 1 mm concentration of gallic acid totally inhibited angiogenesis. In the form of leaf extract, a one-tenth concentration produced the same total inhibition as pure gallic acid. The 10-fold difference in potency suggests the presence of other active compounds contributing to the overall antiangiogenic effect of the extract. The oral absorption of this extract was tested by using serum from rats given the extract orally (gavage) in the angiogenesis assay system. The serum of rats orally administered the sweet leaf tea extract at doses of 0.1% w/w and 0.3% w/w did not significantly inhibit angiogenesis. However, the serum of rats injected intraperitoneally at a dose of 0.1% w/w caused a 41% inhibition of angiogenesis compared with saline injected controls. These preliminary results warrant further bioassay directed identification of other responsible compounds.

Absorption↗
PubMed2006

Development of a human cardiac tissue-based angiogenesis model.

BACKGROUND: Previous angiogenesis models use animal tissues such as the chicken chorioallantoic membrane (CAM) or the rabbit cornea. These models may not accurately reflect the mechanisms responsible for human angiogenesis. MATERIALS AND METHODS: We hypothesized that fragments of human myocardial tissue would develop an angiogenic response from the cut edges of vessels contained within the tissue. To test these hypotheses, we obtained human atrial appendage tissue at the time of cardiac bypass. Fragments of atrial tissue were then incorporated into fibrin thrombin clots. Tissue fragments were observed, and the percent of wells that developed neovessel invasion into the clot was calculated (%I). The subsequent growth of cardiac-derived microvessels was rated and scored over time (Angiogenic Index). RESULTS: There were 20 human atrial appendages plated (n = 24 to 60 wells/specimen) and evaluated in this model. Out of the 20, 16 (80%) atrial appendages developed an angiogenic response in the majority (>50%) of wells plated. Neovessel growth was progressive over 14 to 16 days in culture in all specimens tested. The mean angiogenic index of all specimens was 8.59 +/- 0.91. CONCLUSIONS: This human cardiac tissue-based assay might be useful to screen compounds designed for use in human trials or provide highly vascularized cardiac tissue for autotransplantation. Additionally, the assay provides the foundation to study an individual patent's cardiac tissue and its response to angiogenesis stimulators or inhibitors. This may allow the development of patient-specific therapies designed to enhance revascularization or repair of injured cardiac muscle.

Cell Division↗
PubMed2006

Role of somatostatin analogs in the clinical management of non-neuroendocrine solid tumors.

The somatostatin analogs octreotide, lanreotide and RC-160 (vapreotide) are known to have direct and indirect antitumor effects. Direct effects include the arrest of tumor growth and stimulation of apoptosis, resulting in tumor shrinkage. Indirect antiproliferative effects may occur through antiangiogenesis, immunomodulatory effects and the suppression of tumor-stimulating growth factors. With a safety profile of somatostatin analogs established over 20 years of clinical use in the treatment of neuroendocrine tumors, somatostatin analogs are attractive therapeutic options for patients with non-neuroendocrine tumors. In early clinical trials of somatostatin analogs, however, some cancer patients responded well, while others showed a lack of benefit. This variability in clinical response may reflect the selective binding affinities of octreotide, lanreotide and RC-160, which bind with high affinity to just two of the five different somatostatin receptor subtypes. Treatment response may therefore depend on the specific receptor subtype(s) present in the tumor, the relative proportion of receptor(s) expressed on the tumor cell surface and the absolute quantity of each receptor subtype. Greater understanding of the role of somatostatin receptors, their binding affinities and modes of action has led to increased research into the use of somatostatin analogs, particularly octreotide, in cancer treatment as monotherapies, in combination with hormonal treatments and cytotoxic therapies, and in both adjuvant and neoadjuvant settings. A review of the literature suggests that the antitumor potential of somatostatin analogs should be investigated further and additional studies might determine how these analogs can best be used to improve the treatment of patients with non-neuroendocrine tumors.

Humans↗
PubMed2006

Validation of serum versus plasma measurements of chromogranin a levels in patients with carcinoid tumors: lack of correlation between absolute chromogranin a levels and symptom frequency.

OBJECTIVE: Chromogranin A (CGA) levels are used to confirm the diagnosis and monitor the course of patients with neuroendocrine tumors. Chromogranin A levels are significantly reduced when patients are acutely treated with octreotide; however, limited data are available that correlates octreotide long-acting repeatable (LAR) dose or steady state octreotide blood levels to the absolute value of serum or plasma CGA. METHODS: Plasma, serum, and clinical information on carcinoid syndrome symptoms were collected anonymously from 40 patients treated with long-term octreotide LAR therapy for carcinoid syndrome. RESULTS: We found a strong positive linear relationship exists between serum and plasma CGA levels (r = 0.9858, P < 0.0001). No correlation existed between plasma octreotide levels or LAR dose and the static, absolute plasma/serum CGA levels. Although, higher mean CGA values were seen in the group whose diarrhea was "not under optimal control" than for the group "under optimal control," these results did not reach statistical significance (P = 0.24). Contrary to our hypotheses, a statistically significant inverse relationship was found between the frequency of flushing and the CGA levels (P = 0.0372). Higher mean CGA values were observed in the "under optimal control" group with flushing symptoms. CONCLUSIONS: Either serum or plasma can be used to measure CGA levels. Absolute (static) CGA levels do not positively correlate with symptom intensity during LAR therapy. Dynamic (serial) measurements of CGA are necessary to monitor the effectiveness of medical or surgical therapy.

Antineoplastic Agents, Hormonal↗
PubMed2006

In vitro modeling of the clinical interactions between octreotide and 111In-pentetreotide: is there evidence of somatostatin receptor downregulation?

UNLABELLED: Some authors have suggested that chronic octreotide use enhances the efficiency of radiolabeled somatostatin receptor (sst) imaging. Conversely, desensitization of sst on tumor tissue (tachyphylaxis) may occur occasionally in patients on chronic octreotide therapy. Assuming that chronic exposure to octreotide induces tachyphylaxis, we hypothesized that chronic exposure of sst subtype 2 (sst2)-expressing cells to octreotide would downregulate binding of 111In-pentetreotide to sst and that this downregulation would be due to a reduction in the gene copy number for sst2. METHODS: The clinical scenarios of acute (24 h) and chronic (2 wk) octreotide use, followed by either nuclear imaging exposure (8.6 pmol/L) or therapeutic exposure (510 pmol/L) to (111)In-pentetreotide, were modeled in vitro. Receptor binding in IMR-32 human neuroblastoma cells (high sst2 expression) and PANC-1 human pancreatic cancer cells (no detectable sst2 expression) was evaluated. Gene copy numbers for sst subtypes 1-5 in IMR-32 cells were determined by quantitative polymerase chain reaction. RESULTS: Acute or chronic octreotide exposure at low or high doses did not significantly alter sst2 gene copy numbers or binding of either the diagnostic dose or the therapeutic dose of 111In-pentetreotide. CONCLUSION: In vitro exposure of cells to low or high doses of octreotide for 1-14 d does not result in the development of either tachyphylaxis or upregulation of sst as assessed by changes in gene expression or in high-affinity binding.

Cell Line, Tumor↗
PubMed2005

Black raspberry extract and fractions contain angiogenesis inhibitors.

Targeted therapies, such as agents that inhibit angiogenesis, offer hope as complementary agents in cancer therapy. Angiogenesis-inhibiting agents have the potential for inhibiting tumor growth and limiting the dissemination of metastasis, thus keeping cancers in a static growth state for prolonged periods. Black raspberry (Rubus occidentalis) extract was discovered to be antiangiogenic (0.1% w/v) in a novel human tissue-based in vitro fibrin clot angiogenesis assay. Assay-guided fractionation of a crude black raspberry extract resulted in a highly potent antiangiogenic fraction that accounted for only 1% of the fresh weight of whole black raspberries. At 0.075% (w/v), the active fraction completely inhibited angiogenic initiation and angiogenic vessel growth. Further subfractionation of this active fraction revealed the coexistence of multiple antiangiogenic compounds, one of which has been identified as gallic acid. However, the individual subfractions did not outperform the active whole fraction. These findings suggest that an active black raspberry fraction may be a promising complementary cancer therapy. It is natural and potent enough for manageable dosing regimens. These extracts contain multiple active ingredients that may be additive or synergistic in their antiangiogenic effects. These observations warrant further investigations in animals and human trials.

Angiogenesis Inhibitors↗
PubMed2005

Effect of octreotide LAR dose and weight on octreotide blood levels in patients with neuroendocrine tumors.

OBJECTIVES: Octreotide long acting repeatable (LAR) is widely used for the control of symptoms of functional neuroendocrine tumors. At doses of 30 mg/mo, up to 40% of patients require subcutaneous octreotide "rescue" and up to 40% of patients are given more than 30 mg of LAR/mo. Octreotide acetate binds to the sst2 receptor with an affinity (Kd) of approximately 1 x 10(-9) mol/L (approximately equal to 1000 pg/mL), but higher (approximately equal to 10,000 pg/mL) concentrations of octreotide are required to completely saturate this receptor. Octreotide blood level measurement may be useful to guide LAR therapy in symptomatic patients or in patients who have tumor growth on traditional LAR doses. We hypothesize that LAR doses of 60 mg/mo will produce blood levels of 10,000 pg/mL or greater. At identical monthly LAR doses, patients with higher weights will require more medication to achieve similar plasma octreotide levels than individuals with lower body weights. METHODS: Trough plasma, serum, urine, and saliva octreotide levels were obtained from 52 patients with carcinoid syndrome receiving 20 (n = 8), 30 (n = 19), or 60 mg LAR/mo (n = 10). Octreotide levels were determined by radioimmunoassay. RESULTS: The mean +/- SD plasma octreotide levels for patients receiving 20, 30, or 60 mg LAR/mo were 2518 +/- 1020, 5241 +/- 3004, and 10,925 +/- 5330 pg/mL, respectively. Patient weight (kilograms) was inversely related to plasma octreotide levels. There was a significant correlation between plasma octreotide levels and octreotide levels measured in urine, saliva, and serum. CONCLUSIONS: Frequent measurement of octreotide levels may be useful to guide octreotide therapy in patients with poorly controlled symptoms or those patients experiencing tumor growth.

Adult↗
PubMed2004

The effect of drug dose and drug exposure time on the binding, internalization, and cytotoxicity of radiolabeled somatostatin analogs.

BACKGROUND: Creation of protease-resistant somatostatin analogs has allowed development of these peptides as clinically useful drugs. Widespread diagnostic use of radiolabeled somatostatin analogs has enhanced interest in the binding and intracellular distribution of these peptides. The degree of drug internalization and length of drug retention may be critical for drug-induced cytotoxicity. We hypothesized that the ability of a radiolabeled peptide to bind to a cell, be internalized, and induce cytotoxicity is proportional to both the radioligand concentration and the exposure time. MATERIALS AND METHODS: To test this hypothesis, somatostatin receptor-expressing cells (IMR-32) were incubated with (111)In-pentetreotide, a sst 2 preferring somatostatin analogue. Radioligand exposure time and/or concentration were varied. RESULTS: Prolonged exposure to a fixed concentration of radioligand resulted in progressive increases in whole cell binding and internalization over time. Cells exposed to a relatively fixed number of microCi-Hr yielded constant whole cell binding and internalization. Increasing the microCi-Hr resulted in a proportionate increase in binding. Cytotoxicity was also proportional to the dose of radiation regardless of whether the exposure was internalized radiation (microCi-Hr from (111)In-pentetreotide) or from external beam radiation (cGy). CONCLUSION: Both drug exposure time and drug concentration contribute to cell binding and cytotoxicity in this model and their relative contributions are inversely related.

Biological Transport↗
PubMed2004

A new in vitro assay for human tumor angiogenesis: three-dimensional human tumor angiogenesis assay.

BACKGROUND: A human tissue-based angiogenesis assay is needed to study the biology of angiogenesis in human tumor tissue and to tailor drug selection for patients. METHODS: Fragments of tumor tissue are embedded in fibrin gels containing medium 199, endothelial growth medium, fetal bovine serum, and epsilon-aminocaproic acid. Tumor implants sprout angiogenic vessels that progressively grow into the fibrin matrix. The differential growth pattern of tumor cells and angiogenic vessels in the fibrin gel matrix separates the angiogenic vessels and the tumor stroma into independently observable regions (vessel and tumor compartments). The reproducibility of the assay was tested by using fresh tissue obtained from human tumor xenografts (IMR-32 [neuroblastoma], MDA-MB-231 [breast cancer], and LNCaP [prostate cancer]) grown in nude mice and from fresh surgical breast and thyroid cancer specimens. RESULTS: All tumor fragments studied showed angiogenic sprouting into the fibrin matrix. This created an angiogenic vessel compartment, which was separate from the tumor fragment. The capillary nature of sprouting was confirmed histologically by factor VIII immunohistochemistry. The angiogenic growth fraction was >80% in all groups studied. CONCLUSIONS: This assay may allow functional assessment of the angiogenic potential of human tumors and simultaneous evaluation of a therapeutic agent's antitumor and antiangiogenic effects by virtue of its dual-compartmental structure.

Angiogenesis Inhibitors↗
PubMed2003

A multi-tyrosinated sst1/2 receptor preferring somatostatin agonist inhibits reflex and immune-mediated secretion in the guinea pig colon.

Somatostatin and its analogs such as WOC 3B were compared for their ability to alter the release of 5-hydroxytryptamine (5-HT) and prostaglandins and to affect chloride secretory capacity, determined by activity of neural reflexes or by the influence of immune mediators and other secretagogues. In guinea pig colon set up in flux chambers, the multi-tyrosinated sst1/sst2 receptor preferring somatostatin agonist, WOC 3B, inhibited stroking-evoked 5-HT release without affecting basal release. WOC 3B had no effect on stroking-induced or basal prostaglandin E2 release (PGE2). Neither 5-HT nor PGE2 release was dependent on neural input. Tetrodotoxin induced a decrease in basal short circuit current (Isc) indicative of a decrease in chloride secretion. The decrease in basal Isc during neural blockade was highly correlated with the decrease in basal Isc when WOC 3B was used. In piroxicam- and atropine-treated tissues, to eliminate prostaglandins and cholinergic muscarinic input to crypts, WOC 3B further reduced the piroxicam-resistant and not the atropine resistant Isc during brush stroking the mucosa. Somatostatin and WOC 3B reduced the stroking-evoked Isc with similar half maximum concentrations of 1-2 nM. WOC 3B reduced by more than 50% dimaprit-evoked cyclical Isc. The rank order of potencies in inhibiting dimaprit-evoked Isc was: Somatostatin-14=WOC 3B>CH275=DC-32-92>DC-23-48>> >>DC-32-87=DC-32-97. Low nanomolar concentrations of WOC 3B primarily inhibited the neural effects of carbachol and forskolin on Isc without altering their epithelial effects. Equi-molar concentrations (4 nM) of CH275, a somatostatin sst1 receptor agonist, and the somatostatin sst2 receptor agonist, [Tyr(3)]-octreotide, inhibited dimaprit-evoked Isc by 25% and 26%, and their effects were additive. The results suggest that WOC 3B, a somatostatin analogue containing three tyrosine residues, has anti-secretory effects due to activation of somatostatin sst1 and sst2 receptors on enteric neurons.

Animals↗
PubMed2003

Inhibition of angiogenic initiation and disruption of newly established human vascular networks by juice from Morinda citrifolia (noni).

noni, the juice of the fruit from the Morinda citrifolia plant, has been used for centuries as a medicinal agent. We tested the effects of noni juice in a three-dimensional fibrin clot matrix model using human placental vein and human breast tumor explants as sources for angiogenic vessel development. Noni in concentrations of 5% (vol/vol) or greater was highly effective in inhibiting the initiation of new vessel sprouts from placental vein explants, compared with initiation in control explants in media supplemented with an equivalent amount of saline. These concentrations of noni were also effective in reducing the growth rate and proliferation of newly developing capillary sprouts. When used at a concentration of 10% in growth media, noni was able to induce vessel degeneration and apoptosis in wells with established capillary networks within a few days of its application. We also found that 10% noni juice in media was an effective inhibitor of capillary initiation in explants from human breast tumors. In tumor explants which did show capillary sprouting, the vessels rapidly degenerated (2-3 days) in those exposed to media supplemented with 10% noni.

Angiogenesis Inhibitors↗
PubMed2003

Effect of human Angiostatin protein on human angiogenesis in vitro.

Angiostatin, a 38-kD fragment of plasminogen, inhibits angiogenesis in both animal tumor models and in vitro endothelial cell models. However, human Angiostatin has not been tested in vitro against an intact human tissue target to determine its ability to inhibit the initiation or subsequent promotion of the human angiogenic response. We hypothesized that high doses of human Angiostatin would inhibit the development of an angiogenic response in an intact human vessel target, and would suppress the subsequent growth of blood vessels following the initiation of an angiogenic response. To test these hypotheses, full-thickness human placental vein disks were cultured for 15 days in an in vitro fibrin-thrombin clot assay. This assay system had been used to evaluate the efficacy of a wide variety of compounds. Vessels were obtained from three placentas. Treatments included a control medium plus fetal bovine serum (FBS), heparin-steroid (300 micro g/ml heparin and 350 micro g/ml hydrocortisone; a treatment known to inhibit angiogenesis) and Angiostatin at doses from 1 x 10(-4) to 1 x 10(-9) M. In the control groups, 81% of vessels initiated an angiogenic response compared to 53% of the vessels treated with heparin-steroid. Angiostatin (10(-4)-10(-9) M) decreased the initiation of an angiogenic response, but this was not statistically significant. Of the disks that initiated an angiogenic response, the mean ( +/- standard error of the mean (SEM)) semi-quantitative visual angiogenic index (AI) of the control vessels was 9 +/- 1.7 on day 15. In comparison, the mean AI of heparin-steroid treated vessels was 3.7 +/- 0.4. Angiostatin at doses of 10(-4)-10(-9) M also failed to inhibit blood vessel growth after initiation of the angiogenic response. Based on these observations, we cannot demonstrate significant activity of human Angiostatin (10(-4)-10(-9) M) against the initiation or promotion of a human angiogenic response in this in vitro model of angiogenesis using an intact human vessel target.

Angiogenesis Inhibitors↗
PubMed2003

Development of targeted somatostatin-based antiangiogenic therapy: a review and future perspectives.

Angiogenesis, the development of new blood vessels, is a critical determinant of tumor growth and the dissemination of metastasis. A number of antiangiogenic therapies have been introduced into clinical trials, though few of these are targeted therapies. Somatostatin analogs may be an excellent candidate to develop as targeted antiangiogenic agents alone, or in combination with cytotoxic or cytostatic compounds. Somatostatin analog inhibition of angiogenesis has been demonstrated in the chicken chorioallantoic membrane (CAM) model, the human umbilical vein endothelial cell (HUVEC) proliferation model, and the human placental vein angiogenesis model (HPVAM). This inhibition appears to be the result of a unique upregulation of somatostatin receptor subtype 2 (sst 2) during the angiogenic switch from resting to proliferating endothelium. The distinct overexpression of this receptor provides a unique target for these somatostatin analogs or somatostatin analog conjugates. This manuscript reviews the development of somatostatin analogs as antiangiogenics in both their unlabeled and radiolabeled forms and postulates on future developments in this field.

Angiogenesis Inhibitors↗
PubMed2003

Development of a novel in vitro human tissue-based angiogenesis assay to evaluate the effect of antiangiogenic drugs.

OBJECTIVE: To describe a novel in vitro human tissue-based angiogenic model that can predict an individual tumor's response to antiangiogenic drugs. SUMMARY BACKGROUND DATA: A number of in vitro and in vivo angiogenesis assays exist, but they do not provide potentially useful information for the treatment of an individual patient. Clonogenic assays have been used to evaluate the response of an individual's tumor to antineoplastic agents, but these tumor fragments are cultured in an environment that does not lead to neovessel growth. The authors have previously demonstrated that human vein disks or human tumor xenograft fragments incorporated into a 0.3% fibrin-thrombin clot will develop angiogenic vessel growth from the cut edge of the vessel disk or xenograft fragment. METHODS: Fresh human tumor or normal tissue disks (2 x 1 mm) from fresh surgical specimens were incorporated into fibrin-thrombin clots overlain with nutrient medium containing either 20% fetal bovine serum alone or in combination with Epothilone B, a tubulin inhibitor with antiangiogenic properties. Tissue disks were visually assessed over time to determine the percentage of wells that developed an angiogenic response. Neovessel growth, density, and length were graded at intervals using a semiquantitative visual neovessel growth-rating scheme (angiogenic index, 0-16 scale) devised in the authors' laboratory. RESULTS: Epothilone B treatment at doses of 10-6 mol/L and 10-8 mol/L decreased the number of wells that developed an invasive angiogenic response and limited the development of vessels that invaded the matrix. At these doses, Epothilone B also caused regression of vessels in wells that had been allowed to develop an angiogenic response. Treatment of tumors or normal tissues with Epothilone B at doses less than 10-8 mol/L was ineffective. CONCLUSIONS: Epothilone B may be an effective antiangiogenic agent in a variety of tumor types. The authors speculate that this in vitro model might provide useful information to the clinician on the effect of specific antiangiogenic agents on individual tumors. This may be particularly useful in patients with tumors that, as a group, are unresponsive to treatment with antineoplastic agents.

Angiogenesis Inhibitors↗
PubMed2002

Sentinel nodal assessment in patients with carcinoma of the lung.

BACKGROUND: Assessment of sentinel nodes to predict metastases in a regional nodal basin is valuable for staging patients with melanoma and breast carcinoma. This study tested whether injection of isosulfan blue and technetium-99 could identify mediastinal sentinel nodes in patients with lung carcinoma and determine whether sentinel node histology predicts distal nodal metastases. METHODS: Isosulfan blue and technetium-99 were injected into the tumor and pulmonary resection performed. The hilum and mediastinum were assessed visually and with the gamma probe, and a mediastinal nodal dissection was performed. RESULTS: Thirty-one patients were evaluated. Three patients had positive sentinel nodes and positive distal mediastinal nodes. Twenty-two patients had negative sentinel nodes and negative distal nodes. No sentinel node was identified in 6 patients and 2 patients had two sentinel nodes. CONCLUSIONS: These data demonstrate that this rapid, simple technique can identify sentinel nodes in the mediastinum and that the sentinel node is an accurate predictor of distal nodal metastases in patients with lung cancer.

Aged↗
PubMed2002

Effect of human recombinant Endostatin protein on human angiogenesis.

Tumor growth and metastasis are dependent on the development of new blood vessels. Inhibitors of new vessel growth have been widely investigated as anti-tumor agents. Endostatin, a 20 kDa C-terminal fragment of collagen XVIII inhibits endothelial cell proliferation, induces endothelial cell apoptosis, and can both inhibit and reverse tumor growth in mice. However, human recombinant endostatin has had limited testing against human tissue targets. To investigate the effect of human endostatin on a human vessel target over a broad range of concentrations (10(-l2)-10(-4) M), human placental vein disks were grown for a period of 2 weeks in a 0.3% fibrin clot overlayed with growth medium. Disks from five individual placentas were tested. For each placenta utilized, a control (medium and 20% fetal bovine serum [FBS]) group and a group treated with heparin (300 microg/ml) and hydrocortisone 21-phosphate (350 microg/ml) (heparin-steroid) at a dose known to inhibit angiogenesis were included. Endostatin was tested at concentrations of 10(-12)-10(-4) M in medium containing 20% FBS. The rate of initiation and the angiogenic growth index (on a visually graded semi-quantitative scale of 0-16) were determined for all experimental conditions. Endostatin inhibited angiogenesis in our model only in high concentrations. At 10(-5) M, endostatin did not alter the percent of wells that initiated an angiogenic response, but significantly inhibited subsequent vessel growth. At 10(-4) M, endostatin was able to inhibit both initiation and subsequent new vessel growth. Human endostatin can inhibit the initiation of a human angiogenic response and inhibit the subsequent proliferation of human neovessels when used at high doses in a continuous exposure model.

Angiogenesis Inhibitors↗