Gelfoam Sponge Research Articles

Improved heart function with myogenesis and angiogenesis after autologous porcine bone marrow stromal cell transplantation.

The study evaluated the utility of transplanting bone marrow stromal cells in a porcine myocardial infarction model. A myocardial infarction was created by occluding the distal left anterior descending artery in pigs with coils and Gelfoam sponge. Sternal bone marrow was aspirated, and stromal cells were cultured and induced to differentiate to a myogenic phenotype with 5-azacytidine. Four weeks after coronary artery occlusion, sestamibi technetium single-photon emission computed tomographic scans were performed, and then either a graft of 100 x 10(6) bone marrow stromal cells (n = 5, 30% labeled with bromodeoxyuridine) or culture medium (n = 6) was injected into the infarct region. Four weeks later the tomographic scans were repeated and cardiac function was assessed with pressure and volume measurements. Morphologic and histologic characteristics of the heart were also studied. Histologic examination found bromodeoxyuridine-labeled cells within the infarct region in islands that had sarcomeres and Z-bands and stained positively for cardiac specific troponin I. The bone marrow stromal cell transplant sites had a greater (P <.05) capillary density than did the control sites. The tomographic scans showed that the hearts with the cell transplants had increases in stroke volume, regional perfusion, and wall motion (P <.05 for all groups) relative to the control hearts. The pressure-volume analysis showed improvement (P <.05) in end-systolic elastance and preload recruitable stroke work in the transplantation group relative to the control group. The left ventricular chamber size was smaller (P <.05) and the scar thickness was greater (P <.05) in the hearts with transplanted cells than in the control hearts (P =.06). 5-Azacytidine-treated bone marrow stromal cells transplanted into the myocardial infarct region formed islands of cardiac-like tissue, induced angiogenesis, prevented thinning and dilatation of the infarct region, and improved regional and global contractile function.

Administration of a liposomal FGF-2 peptide vaccine leads to abrogation of FGF-2-mediated angiogenesis and tumor development

Basic fibroblast growth factor (FGF-2) is an important stimulator of angiogenesis that has been implicated in neoplastic progression. Attempts to neutralize or modulate FGF-2 have met with some success in controlling neovascularity and tumor growth. In the present study, two peptides: one corresponding to the heparin binding domain and the other to the receptor binding domain of FGF-2, exerted dose-dependent inhibition of FGF-2-stimulated human umbilical vein endothelial cell proliferation (IC 50=70 and 20 μg/ml, respectively). The identification of these functional regions suggested that targeting these domains might be an approach for the modulation of FGF-2 function. To investigate this possibility, we vaccinated mice with either the heparin binding domain peptide or the receptor binding domain peptide of FGF-2 in a liposome/adjuvant format, and analyzed the effect of vaccination on FGF-2-driven angiogenesis, tumor development and immune status. Mice vaccinated with the heparin binding domain peptide generated a specific antibody response to FGF-2, blocked neovascularization in a gelfoam sponge model of angiogenesis, and inhibited experimental metastasis by >90% in two tumor models: the B16BL6 melanoma and the Lewis lung carcinoma. These effects were not observed in mice treated with the receptor binding domain peptide conjugated to liposomes or liposomes lacking conjugated peptide. These data suggest that a heparin binding domain peptide of FGF-2, when presented to a host in a liposomal adjuvant formulation, can ultimately lead to inhibition of angiogenesis and tumor growth.

Bio-stretch, a computerized cell strain apparatus for three-dimensional organotypic cultures.

In the present study, a unique mechanical strain apparatus for three-dimensional organotypic cultures was developed into a computerized system. It consists of a personal computer running Windows-based software, the Bio-Stretch Manager, a Bio-Stretch Controller, and three sets of magnet boards. Cells are cultured on a Gelfoam sponge that is placed in a 35 mm petri dish with one end glued to the dish, and the other end attached to a coated steel bar. The petri dish is placed in front of a magnet, and the movement of the steel bar is controlled by dynamically changing the magnetic field. Up to five stretch patterns of variable frequency, duty cycle, and magnitude can be designed for each stretch regimen. Three different stretch regimens can be tested simultaneously. The operational characteristics of sponges were examined. Attachment of cells to the sponges was observed on several cell types. These features provide wide options for using this system to study the effects of mechanical stretch on cells.

Partial splenic embolization for hypersplenism concomitant with or after arterial embolization of hepatocellular carcinoma in 30 patients

To study the value of partial splenic embolization (PSE) for the treatment of hypersplenism in patients undergoing embolization of hepatocellular carcinoma (HCC). Transcatheter hepatic arterial embolization (THAE) combined with PSE was performed in 30 patients with HCC complicating liver cirrhosis, portal hypertension, and hypersplenism. Gelfoam sponge was used as the embolic material for PSE and limited to 100-150 pieces. More than 50% of splenic parenchyma was infarcted in 27 patients. Leukopenia and thrombocytopenia were corrected by PSE in 25 of 27 patients with hypersplenism. In 26 patients with esophageal varices, including 5 patients with bleeding, no rebleeding occurred during a 6-17 month follow-up. Hypersplenism was not corrected in 2 of 3 patients whose infarcted splenic parenchyma was less than 50%. No splenic abscesses or other severe complications were observed. Of the 30 patients treated, 19 are still alive after 1 year. THAE combined with PSE is a safe and effective measure for patients with HCC.

Vitamin A-soaked Gelfoam Sponges and Wound Healing in Steroid-treated Animals

Previous work has shown improved wound healing after the administration of systemic vitamin A in patients on chronic steroids. In contrast there have been mixed reports on the effect of topical vitamin A on wound healing in steroid-treated patients. Previous laboratory work has suggested that the topical application of vitamin A may be beneficial to wound healing in a sutured wound in a steroid-treated rat. Due to some inconsistencies in previous studies and steroid animal models, we sought to develop a better wound-healing model in a steroid-treated rat and to assess the effect of topical vitamin A as part of the wound closure. With preliminary studies, we developed a consistent and reliable wound-healing model in a steroid-treated rat using dexamethasone in contrast to cortisone acetate, which had been used in previous studies. Next, rats were randomized into 8 groups, some of which received steroids. Wounds were treated with saline or vitamin A topically or via a soaked gelfoam sponge. Rats were wounded 1 week after the commencement of steroid administration. Wounds were repaired and allowed to heal for 2 weeks. Strips of the wounds were then harvested and tested for tensile strength and breaking strength using a tensiometer. Wound edges were then fixed and wound surface area was measured using digital planimetry. Steroid treatment resulted in consistent weight loss and failure to gain weight as well as decreased breaking strength. Tensile strength was not decreased. Vitamin A applied for 10 minutes before wound closure and a gelfoam sponge alone placed before wound closure both resulted in an increased breaking strength and tensile strength.(ABSTRACT TRUNCATED AT 250 WORDS)

Epithelial tissue formation in matrix culture and in histophysiologic gradient culture

Formation of epithelial tissues in culture so that they become facsimiles in their structure of such tissues in nature requires procedures that comply with several spatial imperatives: a) three-dimensional growth; b) histophysiologic conditions that provide, concurrently, gradients of maturation and of diffusion of metabolites; and c) growth as layers of cells without free edges. Many steps have been required in the evolution of these methods. Two systems are described here in sufficient detail to serve as a manual. Three-dimensional growth of masses of epithelial tissue is accomplished in matrix culture using Gelfoam sponge and collagen-coated cellulose sponge. Radial gradient culture, a recent development, provides conditions that comply with the requirements of histophysiologic gradients and of epithelial tissue growth in layers without interruption in their continuity.

Short-term and long-term results of experimental wrapping techniques for bronchial anastomosis

Major complications of bronchial anastomoses for either transplantation or sleeve resection include early leak, fistula formation, granulation tissue, and stenosis. To evaluate the impact of technique on these complications, we designed a nonimmunocompromised canine model with a totally ischemic bronchial segment. We wished to discover the prevalence of early and late complications of a telescoping anastomosis and if wrapping techniques modify them. We autotransplanted 25 mm of left main-stem bronchus by telescoping 1 mm of proximal into distal bronchus sutured with interrupted 4-0 polyglactin sutures. The animals were divided into four groups: no wrap; omental pedicle wrap; detached-free omental wrap; and Gelfoam sponge soaked in porcine omental extract. Weekly bronchoscopic studies assessed airway patency. After the animals were put to death at 70 days, the luminal areas of the proximal anastomoses were compared with the origin of the left main bronchus. No air leak, bronchial disruption, or infection occurred in any group at any time. Luminal narrowing occurred in all four groups but was most pronounced in the three groups in which wrapping techniques were used. We conclude that wrapping of a telescoped anastomosis is not necessary to prevent early complications. However, no method completely eliminates stenosis. Further experiments should determine the effects of immunomodulation in this model.

Recurrent hepatocellular carcinoma after partial hepatectomy: value of treatment with transcatheter arterial chemoembolization.

The usefulness of transcatheter arterial chemoembolization (TACE) of the hepatic artery was retrospectively evaluated in 66 patients who underwent the procedure for treatment of hepatocellular carcinoma that recurred after partial hepatectomy. The materials infused were Gelfoam sponge or Gelfoam sponge plus Lipiodol and an anticancer agent. A control group of 15 patients with recurrent tumor received oral anticancer agents alone. The cumulative survival rate for the TACE group was 88% for the first year, 57% for 2 years, 42% for 3 years, and 27% for 5 years, whereas that of the control group was 80% for the first year, 27% for 2 years, and 18% for 3 years. Thus, the prognosis of the TACE group was significantly better (p less than or equal to .01, log-rank test) than that of the control group. The survival rate was inversely correlated with the ratio of the volume of the recurrent tumor to the volume of the whole residual liver. These results suggest that TACE is more effective than oral chemotherapy for treatment of hepatocellular carcinoma that recurs after partial hepatectomy.

Relationship of Invasiveness to Proliferating Activity and to Cytoskeletal Protein Production in Human Neuroepithelial Tumors Maintained in an Organ Culture System: Use of Human Cortex and Dura as Supporting Matrices

Abstract The proliferation and invasiveness of cultured human neuroepithelial tumors were studied. A human malignant astrocytic glioma cell line (U-251 MG) and a medulloblastoma cell line (D283 Med) were maintained for 3 weeks in an organ culture system using adult human brain cortex, dura mater, or Gelfoam sponge as growth matrices. The cells were labeled with bromodeoxyuridine (BrdU) at different time points, and immunohistochemistry was performed for BrdU, glial fibrillary acidic (GFA) protein (in U-251 MG), and neurofilament (NF) protein (in D283 Med). In the U251 MG line, the cells grew successfully in each matrix, forming a fibrillated solid area and a peripheral zone of invasion. The labeling index (LI) expressed as the percentage of BrdU-labeled cells and the percentage of GFA protein-positive cells in the two zones of the explants were analyzed. The LIs in all cultures were significantly higher in the peripheral than in the central zones. On the other hand, the percentage of GFA protein-positive cells in each matrix was greater in the central zone than in the periphery. The LI was inversely correlated with the percentage of GFA protein-positive cells over the areas counted in each growth matrix. GFA protein production in cells grown on cortex and on dura mater was significantly higher than that in cells grown on Gelfoam. In the D283 Med line, the cells formed an aggregated zone, with peripheral cells infiltrating the Gelfoam. This line showed poor growth on human cortex. Cells grown on the dura demonstrated an LI similar to that on Gelfoam, and cells often infiltrated the dura. The LI of these cells was higher in the peripheral than in the central zone of the explants, both on dura and on Gelfoam. The cells showed variable immunopositivity for each NF protein subunit. No apparent difference in the percentage of immunopositive cells for each NF protein subunit was detected between cells grown on dura and on Gelfoam; however, cells grown on dura often demonstrated neurite-like processes. The results indicate that an organ culture system using human brain cortex or dura mater as matrices for in vitro growth is a promising tool for the study of neuroglial differentiation and of the invasiveness of cultured human neuroepithelial tumors.

Relationship of invasiveness to proliferating activity and to cytoskeletal protein production in human neuroepithelial tumors maintained in an organ culture system

The proliferation and invasiveness of cultured human neuroepithelial tumors were studied. A human malignant astrocytic glioma cell line (U-251 MG) and a medulloblastoma cell line (D283 Med) were maintained for 3 weeks in an organ culture system using adult human brain cortex, dura mater, or Gelfoam sponge as growth matrices. The cells were labeled with bromodeoxyuridine (BrdU) at different time points, and immunohistochemistry was performed for BrdU, glial fibrillary acidic (GFA) protein (in U-251 MG), and neurofilament (NF) protein (in D283 Med). In the U-251 MG line, the cells grew successfully in each matrix, forming a fibrillated solid area and a peripheral zone of invasion. The labeling index (LI) expressed as the percentage of BrdU-labeled cells and the percentage of GFA protein-positive cells in the two zones of the explants were analyzed. The LIs in all cultures were significantly higher in the peripheral than in the central zones. On the other hand, the percentage of GFA protein-positive cells in each matrix was greater in the central zone than in the periphery. The LI was inversely correlated with the percentage of GFA protein-positive cells over the areas counted in each growth matrix. GFA protein production in cells grown on cortex and on dura mater was significantly higher than that in cells grown on Gelfoam. In the D283 Med line, the cells formed an aggregated zone, with peripheral cells infiltrating the Gelfoam. This line showed poor growth on human cortex. Cells grown on the dura demonstrated an LI similar to that on Gelfoam, and cells often infiltrated the dura.(ABSTRACT TRUNCATED AT 250 WORDS)

A New Hemostatic Procedure for Percutaneous Transhepatic Portal Vein Catheterization

To avoid major complications, such as intraperitoneal bleeding after percutaneous transhepatic catheterization, a new technique has been developed. In this procedure, a stick-like gelfoam sponge compressed manually and loaded in a cartridge, is inserted through an outer sheath catheter after the removal of the catheter, and the puncture hole is plugged. No bleeding has been experienced in 56 patients with various hepatic diseases since using this method.

Effect of a topical drug combination on the early healing of extraction sockets in the vervet monkey

An animal trial was undertaken in 9 vervet monkeys to test the effect of a medicament combination on extraction socket healing. All 4 third molar teeth were extracted and 2 of the sockets in each animal were packed with Gelfoam sponge impregnated with a medicament containing a local anaesthetic, antiseptic, 2 potent antifibrinolytic agents and metronidazole. The remaining 2 sockets acted as controls: one of the sockets was packed with Gelfoam sponge alone and the other allowed to heal spontaneously with no implant. The healing of these sockets was evaluated histometrically in 4-, 6- and 8-day specimens, and from the results it was concluded that sockets containing the medicament compound showed enhanced early socket healing.

Explant organ culture of hamster alimentary tract epithelium.

Adult Syrian Golden hamster alimentary tract maintained as explants in organ culture was studied using the model system for hamster pancreas described by Resau et al. (1983a). Explants of esophagus, stomach, duodenum and colon were maintained in organ culture on Gelfoam sponge rafts in a high-oxygen atmosphere with serum-supplemented CMRL-1066 medium. All of the tissues were observed to show evidence of sublethal acute cell injury during the first several days of culture. Subsequently, the epithelial tissues recovered from this injury, repopulated the denuded areas of the explants and replicated within the sponge matrix. Explants were maintained in a differentiated state for 30+ days and sampled for morphology to examine to process of cell injury, repair, differentiation and replication which occurs in mucosal epithelia. The percentage of basement membrane covered by epithelia in the explants from various tissues was compared to the level of LDH in the media to reveal the relationship between viability determined by biochemical and by morphological methods. Restitution of the mucosal surface occurred in all of the explants. We conclude that adequate populations of replicating cells are maintained within the epithelium of the hamster alimentary tract tissues in vitro so that restitution can occur through migration and subsequent differentiation of the epithelial cells within the mucosa of the explants.

3H]Lysine incorporation into protein of hemisected, cycloheximide-treated spinal cord

A single application of a protein synthesis inhibitor, puromycin, in the lesion site of a spinal hemisection transiently decreases protein incorporation for less than 24 h and results in axonal sprouting and dendritic and synaptic stability for at least 60 days. In this study we characterized the protein-altering properties of single applications of a different protein synthesis inhibitor, cycloheximide, for comparison. The protein incorporation of [ 3H] lysine into hemisected rat spinal cord (left side, T2) treated with cycloheximide (100 μg/ml in Gelfoam sponge placed in wound at time of lesion) was tested 3 h, 6 h, 12 h, 1, 3, 7, and 14 days later and compared with that in 0.9% saline in Gelfoam implants as normal controls ( N = 54). One hour prior to utilization, the animals were injected subcutaneously with 200 μCi l-[4,5(n)- 3H]lysine monohydrochloride. No treatment-related differences in brain radioactivity were noted at any time postoperation. In spinal cord, inhibition of amino acid uptake by cycloheximide was not demonstrable at 3 h after hemisection, but could be detected from 6 h to 1 day after implantation. During that time, the TCA-soluble radioactivity at the lesion site was greater than in control animals ( P < 0.05). These results suggest that as with puromycin, morphologic effects produced by cycloheximide on spinal cord regeneration may correspond to chemical events which occurred within the first 24 h after cord hemisection and cycloheximide treatment.

Organ explant culture of adult Syrian golden hamster pancreas.

An organ explant culture system has been developed for long term maintenance of adult pancreatic tissue from the Syrian golden hamster. Gastric and duodenal lobe explants of up to 0.5 cm2 size were placed in tissue culture dishes (60 mm2) on Gelfoam sponge rafts to which was added 5 ml of CMRL medium 1066 supplemented with heat inactivated newborn bovine serum, L-glutamine, hydrocortisone, insulin, and antibiotics. Dishes were placed in a controlled atmosphere chamber, which was gassed with 45% O2, 50% N2, and 5% CO2 and incubated at 36.5 degrees C. Viability of the tissues was determined by light and electron microscopy as well as by [3]thymidine incorporation. Explants were viable for up to 70 d. Zymogen granule-containing cells characteristic of acinar cells and mucus-containing cells characteristic of ductal cells were present throughout this period. However, endocrine cells were only present for the 1st wk in culture.

DIFFERENTIATION OF A PRIMARY CHEMICALLY INDUCED RAT NEPHROBLASTOMA IN ORGAN CULTURE.

The differentiation in organ culture of a rat nephroblastoma is compared with differentiation of normal rat metanephric tissue under the same conditions. The nephroblastoma arose in a 19 week old female Fischer F344 rat given a single intraperitoneal injection of 4.0 μmole methyl(methoxymethy1)nitrosamine (DMN-OMe)/g body weight at one day of age. The tumor consisted almost entirely of spindle cells although a few well-differentiated tubules were scattered throughout the tumor mass. No primitive tubules were seen, but focal aggregates of tumor cells suggestive of nascent epithelial differentiation were frequent. Fragments of the nephroblastoma were cultured on gelfoam sponge in Williams Medium E supplemented with hydrocortisone, insulin, and fetal bovine serum. Within one day extensive tubulogenesis was observed. High mitotic activity resulted in a steady increase in the size of cultured explants over a period of 6 days. By day six, differentiating tubules filled the explant tissue. Cultured fragments were nearly indistinguishable histologically from normal F344 rat fetal kidney explanted to organ culture on day 15 of gestation and grown in vitro for the same period.

Effect of puromycin administration on presynaptic bouton regeneration after hemisection of rat spinal cord

The effect of puromycin on synaptic renewal on spinal neurons 1 to 1.5 mm rostral to the site of a T1-T2 interface after hemisection was studied. Ninety-five Long-Evans rats were utilized (five animals per group) 10, 20, 30, 45, 60, and 90 days posthemisection. The groups consisted of normal animals, those with spinal cord hemisection alone, those with Gelfoam sponge saturated with 0.9% saline placed in the lesion, or those with a Gelfoam sponge saturated with 1.0 m m puromycin dihydrochloride in 0.9% saline placed in the lesion (at surgery). The spinal cords were processed for the light microscopic impregnation of presynaptic boutons. Counts of boutons (lamina IV and IX motoneurons) were made from coded material and analyzed statistically. After spinal cord hemisection alone, there was a cyclic significant renewal of presynaptic boutons on the somata of lamina IV and IX neurons with decreases ( P < 0.05) at 10 to 20 days, increases ( P < 0.05) at 30 days, and spontaneous loss ( P < 0.05) at 45 days. This small number of boutons was retained at 60 days postoperative (DPO) and remains at this value at 90 days on lamina IV neurons or increases on lamina IX neurons. Saline-puromycin-Gelfoam implantation resulted in a significant bouton decrease ( P < 0.05) on lamina IX neurons at only 45 days postoperative and no differences from normal on lamina IV neurons. These data show a “protective” effect that stabilized presynaptic boutons on lamina IV neurons for 90 days and on lamina IX neurons for at least 30 days.

Effects of puromycin on incorporation of [ 3H]lysine into protein following hemisection of rat spinal cord

Single applications of puromycin into the lesion site of spinal hemisected rats were previously shown to induce transient nerve fiber growth into the lesion. We determined the protein incorporation of [ 3H]lysine into rat spinal cord and brain treated with puromycin in groups of animals 3 h, 12 h, and 1, 3, 7, and 14 days after a left spinal cord hemisection at T2 and compared it to that in saline-treated controls. Each group consisted of two control animals, in which a left spinal hemisection was made at T2 and a Gelfoam sponge soaked in saline implanted in the lesion, and five experimental animals which were implanted with Gelfoam soaked in 1 m m puromycin in saline. One hour prior to utilization, animals were injected subcutaneously with 200 μCi l-[4,5( n)- 3H]lysine monohydrochloride. In brain, no hemispheric or regional differences in protein or soluble fraction radioactivity could be detected in puromycin-treated or control animals. In spinal cord, inhibition of amino acid uptake was not demonstrable 3 h after hemisection, but could be detected at the site of the lesion 6 and 12 h after implantation. Three and 7 days after spinal hemisection, puromycin-treated animals exhibited a larger uptake of amino acid into protein at the lesion site than controls. The results suggest that morphological effects produced by puromycin on spinal cord regeneration may correspond to chemical events occurring within the first 24 h after cord hemisection and puromycin implantation.

Effect of puromycin treatment on the regeneration of hemisected and transected rat spinal cord

The effect of puromycin on spinal cord regeneration was studied following implantation into the site of spinal cord hemi- or transection of Gel-foam saturated with puromycin (1 mM) in a saline carrier, implantation of Gel-foam sponge saturated with saline (carrier control), or lesion alone (lesion control). The spinal cords of 107 rats were studied with light and electron microscopy 7, 14, 30, 60 and 90 days postoperative (DPO). Spinal cord hemisected animals developed a dense cicatrix at the site of lesion replete with connective tissue, blood vessles, and myelinated and unmyelinated nerve fibres which could be traced to peripheral sources. Rostrally at the C.N.S.--cicatrix interface, there were reactive neuroglial cells, occasional nerve fibres and finger-like projections of spinal cord (due to cavitation lesions) which contained neuroglia, axons and dendrites. Implantation of saline in Gel-foam resulted in the same morphology as in hemisected animals except for increased lesion size due to mechanical factors and decreased cicatrix density during the first 30 DPO. Puromycin treatment resulted in a cicatrix with initial decreased cell density but which contained a new class of nerve fibres at 30 DPO. These nerve fibres were oriented in a rostro-caudal direction, were unmyelinated, 0.1-0.2 micron in diameter and had expanded smooth endoplasmic reticulum. Some of these nerve fibres were degenerating at 30 DPO and all were absent by 60 DPO. The puromycin-treated spinal cord within 200 micron rostral to the basal lamina contained nerve terminal conglomerates, which resembled boutons, in fascicles from 30-90 DPO (duration of experiment). Hemisection of the spinal cord by crushing 1-1 1/2 segments rostral to the site of puromycin implantation at 30 DPO resulted in degeneration of these nerve fibres in the cicatrix as well as the degeneration of nerve terminal conglomerates just rostral to the basal lamina. The regenerative capacity of the spinal cord is discussed in relationship to these findings.

Transcatheter therapeutic embolization in the management of massive upper gastrointestinal bleeding.

Two methods of controlling life-threatening hemorrhage are infusion of vasospastic agents and embolization of the bleeding vessel. Gelfoam powder, Gelfoam sponge, and isobutyl cyanoacrylate have been used to control severe arterial and variceal hemorrhage from the upper gastrointestinal tract. The authors summarize their results and discuss gastric necrosis, a serious complication.