Serum IL-15 and IL-15Rα levels are decreased in lean and obese physically active humans.

Circulating IL-15 presence is required to stimulate anti-adipogenic effects of the IL-15/IL-15Rα axis in adipose tissue. Although exercise increases blood IL-15 expression post-exercise, it remains inconclusive whether physical activity can alter the baseline concentrations of this cytokine. The aim of this study was to determine whether physical activity regulates circulating IL-15 and IL-15Rα in lean and obese individuals. Two hundred and seventy-six participants were divided into five groups according to physical activity (PA), body mass and type 2 diabetes mellitus (T2DM) diagnosis: (a) lean PA (N = 25); (b) lean non-PA (N = 28); (c) obese PA (N = 64); (d) obese non-PA (N = 79); and (e) obese non-PA with T2DM (N = 80). Serum IL-15 and IL-15Rα, blood glucose/lipid profile and body composition were measured. Serum IL-15 and IL-15Rα decreased in PA participants compared to non-PA (P < .05), while IL-15 and IL-15Rα increased in obese with T2DM compared to obese without T2DM (P < .05). No differences were observed between lean non-PA and obese PA. Serum IL-15Rα was associated with fasting glucose (R2 = .063), insulin (R2 = .082), HbA1c (R2 = .108), and HOMA (R2 = .057) in obese participants. Circulating IL-15 and IL-15Rα are reduced in lean and obese participants who perform physical activity regularly (≥180 min/week), suggesting a regulative role of physical activity on the circulating concentrations of IL-15 and IL-15Rα at baseline. Moreover, the relationship observed between IL-15Rα and glucose profile may indicate a role of the alpha receptor in glucose metabolism.

The angiogenesis promoter, proadrenomedullin N-terminal 20 peptide (PAMP), improves healing in both normoxic and ischemic wounds either alone or in combination with autologous stem/progenitor cells.

A combination of vascular pathologies and other complicating factors results in chronic wounds which constitute a serious burden for both patients and national health systems, due to prolonged hospital stays, high costs, and prolonged nursing staff dedication. Here we investigate whether proadrenomedullin N-terminal 20 peptide (PAMP), a naturally occurring peptide of the skin with antimicrobial and proangiogenic properties, either alone or in combination with autologous skeletal muscle stem/progenitor cells, acts as a wound healing factor. The rabbit ear was chosen as a test system, since it offers a reliable model for normoxic and ischemic wounds. Topical treatments with PAMP, stem/progenitor cells, or a combination of both, resulted in significant improvements of healing, when compared to untreated wounds. PAMP was very effective in promoting reepithelialization and angiogenesis, whereas treatment with stem/progenitor cells alone resulted in less wound contraction. Interestingly, the combination of PAMP and stem/progenitor cells, while maintaining angiogenic potency, reverted to the contraction levels observed in the untreated controls. Under ischemic conditions, generalized necrosis of the dermis and the underlying cartilage was observed in untreated wounds. Treatments of these wounds with PAMP or stem/progenitor cells allowed a timely recovery. In conclusion, PAMP either alone or in combination with autologous stem/progenitor cells may provide a useful tool for improving wound healing both in normoxic and ischemic conditions.

Immune response to the long-term grafting of cryopreserved small-diameter arterial allografts.

INTRODUCTION: The viability and immunological response induced by cryopreserved arterial allografts remain unclear. This study examines the post-graft behaviour of this type of vessel substitute. MATERIALS AND METHODS: Both iliac arteries were extracted from Lewis rats (donors) and used to establish groups of allogeneic fresh (group I) or cryopreserved (group II) grafts in Fisher-344 rats (recipients). Cryopreserved segments for grafting were prepared by automated controlled freezing at a cooling rate of 1°C/min followed by storage in liquid nitrogen vapour at -145°C for 30 days. Before grafting, the vessels were slowly thawed. Animals were sacrificed at 14, 30, 90 and 180 days post-surgery when graft specimens were obtained for light and electron microscopy and immunohistochemical detection of inflammatory cells (CD45, ED1, CD4, CD8). RESULTS: After surgery, 85.71% of the grafts in group I and 82.14% in group II were patent. Following long-term implant, both the fresh and cryopreserved allografts showed complete loss of the muscle compartment of the media. Inflammatory or CD45-positive cells (mainly macrophages and CD8 T-lymphocytes) were detected at earlier time points in suture zones and adventitia. In the fresh allografts, the number of immunolabelled cells steadily increased until they were seen to occupy the entire adventitia at 90 days, with high numbers persisting at 6 months. In the cryopreserved allografts, this adventitial inflammatory infiltrate was significantly reduced. CONCLUSIONS: The cryopreservation/slow thawing protocol used diminished the immune response induced by fresh arterial allografts improving their behaviour after grafting.

Comparing the behavior of different polypropylene meshes (heavy and lightweight) in an experimental model of ventral hernia repair.

New generation prosthetic biomaterials for abdominal wall repair have been designed to be less dense, by having larger pores than that of the standard polypropylene meshes, to improve abdominal wall compliance. The aim of the present study was to analyze the functional and morphologic properties of these new meshes. For this purpose, 7 x 5 cm(2) defects were created in the anterior abdominal wall of 36 male New Zealand White rabbits and repaired using different polypropylene meshes: a heavyweight mesh (HW), Surgipro, and two lightweight meshes (LW), Parietene and Optilene. Six animals each implanted with biomaterial were sacrificed on postoperative days 14 and 90. Histological and morphometric analysis, adhesion assessment, and biomechanical resistance tests were performed. Similar behavior was shown by the LW and HW meshes in terms of the adhesions and macrophage response induced. After 14 days, the tensile strength of Optilene was greater than the strengths recorded for the other two biomaterials, probably because of its high elasticity. By 90 days, however, the tensile strengths of the three biomaterials were comparable. In conclusion, despite an initial tensile strength advantage shown by the mesh with larger pores, at 90 days postimplant, tensile strengths were similar. Compared with HW, LW prostheses have the benefit that less foreign material was implanted, preserving the elasticity of the recipient host tissue.

Cell viability and prostacyclin release in cultured human umbilical vein endothelial cells.

Construction of efficient substitutes of human blood vessels is strongly dependent on the use of viable and fully functional cultured endothelial cells (ECs). However, very few reports have been published to date focused on the evaluation of cell viability of cultured ECs. In this work, we have determined cell viability, von Willebrand factor, and prostacyclin (PGI(2)) activity in primary cell cultures of human umbilical vein ECs, to identify the specific cell passage that is more appropriate for the development of artificial organs by tissue engineering. Cell viability was determined by quantification of the intracellular concentration of several ions by highly sensitive electron probe X-ray microanalysis, whereas von Willebrand was assayed by immunohistochemistry and PGI(2) release was quantified by radioimmunoassay. The results of our analyses demonstrate that the K/Na ratio was different for each cell passage (4.72 for the first passage, 4.55 for the second passage, and 7.82 for the third passage), suggesting that the highest cell viability corresponds to the third passage. In contrast, PGI(2) production was higher at the first two cell passages, with a significant decrease at the third passage (6.46 +/- 0.10, 5.98 +/- 0.08, and 1.62 +/- 0.05 ng/mL of supernatant for the first, second, and third passages, respectively), whereas von Willebrand expression was similar among the three cell passages analyzed in this work (64.12%, 66.66%, 65.93% of positive cells, respectively). These data suggest that cells corresponding to the second cell passage show the best ratio of viability to functionality and should therefore be used for tissue engineering protocols.

Efficiency of 4% icodextrin in preventing adhesions to spiral tacks used to fix intraperitoneal prostheses.

BACKGROUND/AIMS: This study was designed to evaluate the extent of adhesion formation to prostheses fixed with spiral tacks and to establish whether the use of Ringer's lactate or icodextrin could prevent these adhesions. METHODS: 24 New Zealand white rabbits weighing around 3,000 g were implanted with a 7 x 5 cm patch of ePTFE (DualMesh) through a midline laparotomy. The prosthesis was fixed to the intact peritoneum using spiral tacks. Three study groups were established according to whether the animals were: implanted with ePTFE fixed with spiral tacks or implanted with ePTFE fixed with spiral tacks and simultaneously administered Ringer's lactate or 4% icodextrin in the peritoneal cavity. Adhesion formation and prosthetic behavior at the prosthesis/peritoneal interface were evaluated and quantified by sequential laparoscopy performed at 3, 7 and 14 days. RESULTS: Adhesions generally formed on the tacks and were classified as the fully integrated type. No significant differences were observed in terms of the extent of adhesions or of neoperitoneal thickness between control animals and those receiving Ringer's lactate or icodextrin. CONCLUSIONS: (a) Prosthesis-fixing tacks induced adhesions; (b) the use of substances such as icodextrin or Ringer's lactate does not seem to diminish adhesion formation, and (c) the use of icodextrin offered no benefits over that of Ringer's lactate solution.

Influence of the structure of new generation prostheses on shrinkage after implant in the abdominal wall.

When a biomaterial is used to repair an abdominal wall defect, wound contraction can cause the prosthesis to shrink, and the tension generated can provoke recurrence of the defect. This study was designed to determine whether the structure of a prosthesis can directly influence prosthetic shrinkage. Abdominal wall defects (7 x 5 cm) in rabbits were repaired using the laminar prosthesis DualMesh (DM), the composites Sepramesh (Se) and Vypro II (Vy), and the reticular prosthesis Surgipro (PP). The animals were sacrificed 14 and 90 days after surgery, at which time implant specimens were morphologically and immunohistochemically examined to establish the presence of myofibroblasts and macrophages. The size of each prosthesis was measured at the end of the study through image analysis. Morphometric measurements revealed greatest prosthesis shrinkage for Se, PP, and Vy (16.05% +/- 5.08%; 13.75% +/- 4.22%; 16.16% +/- 6.34%), while the DM prostheses only showed a 7.57% +/- 0.62% decrease in size (p < 0.05). In the DM implants, the macrophage response and myofibroblast labeling were reduced. Both biomaterial structure and the macrophage reaction induced at the implant site modulate prosthetic shrinkage, laminar prostheses of the ePTFE type undergoing less shrinkage than reticular meshes. Reduced DM shrinkage was linked to decreased myofibroblast numbers 2 weeks postimplant.

Composite prostheses for the repair of abdominal wall defects: effect of the structure of the adhesion barrier component.

The component of a composite prosthesis, which makes contact with the visceral peritoneum, can be reabsorbable or non-reabsorbable, and laminar or reticular. This study was designed to determine whether the composition of this second, barrier component could improve its behavior at this interface. Abdominal wall defects in rabbits were repaired using a polypropylene prosthesis (PP), or the composites Sepramesh (PP + h) or Vicryl (PP + v). Fourteen days after surgery, the implants were evaluated by light and scanning electron microscopy, and immunohistochemistry. Prosthetic areas occupied by adhesions (PP: 71.08 +/- 5.09, PP + h: 18.55 +/- 4.96, P + v: 69.69 +/- 16.81%), neoperitoneal thickness (PP: 256.17 +/- 21.68, PP + h: 83.11 +/- 19.63, PP + v:213.72 +/- 35.90 microm) and macrophage counts (PP: 8.73 +/- 1.16, PP + h: 27.33 +/- 4.13, PP + v: 31.24 +/- 3.08%) showed significant differences (P < 0.05). The tested biomaterials induced an optimal recipient tissue infiltration. Least adhesion formation was observed on the PP + h implants. This suggests that the second component, although reabsorbable, should be smooth in structure.

Composite prostheses used to repair abdominal wall defects: physical or chemical adhesion barriers?

In a composite prosthesis, the component placed at the peritoneal interface takes the form of a physical or chemical barrier. In this experimental study performed on the white New Zealand rabbit, several composites were examined to establish the effectiveness of these barriers at impeding adhesion formation. The biomaterials tested were two polypropylene prostheses (PP) with the physical barriers of expanded polytetrafluoroethylene or polyurethane (PP + ePTFE and PP + PU) and two prostheses (one polyester and the other PP) with the absorbable chemical barriers of polyethylene glycol/glycerol and hyaluronate, respectively (PO + gl and PP + hy). The composites were used to repair 7 x 5 cm defects created in the abdominal wall of the animals by placing the implant in contact with the visceral peritoneum and the subcutaneous tissue and fixing it to recipient tissue by 4/0 polypropylene running suture. Fourteen days after surgery the animals were sacrificed and specimens were taken for light microscopy and scanning electron microscopy. Adhesions developing at the prosthesis/visceral peritoneal interface were quantified. All the prostheses induced optimal mesothelialization. Composites with physical barriers behaved similarly in terms of provoking adhesions. However, the prostheses with chemical barriers differed in their effectiveness at preventing adhesions. Overall, the best results were obtained with the PP + PU composite.

Histological evaluation of scar tissue inflammatory response: the role of hGH in diabetic rats.

This paper describes a polymer site-specific delivery system containing human growth hormone in an in vivo model of scarring in the diabetic state. Copolymer discs with the hormone were introduced into incisions made in rats previously injected with streptozotocin in order to induce diabetes. Tissue specimens for evaluation were obtained at 3, 7 or 10 days after the procedure. Study groups were healthy rats and diabetic rats untreated or treated with/without the hormone. Histological sections were prepared for light microscopy examination of wound zones. Three and 7 days after surgery, polymer remains could be observed in the subcutaneous tissue. These remnants induced a moderate foreign body reaction. The number of macrophages detected was directly related to neovessel formation and metalloelastase expression. The CD4+/CD8+ ratio was low during the initial follow up stages (3 and 7 days) in untreated diabetic rats, yet an increased ratio corresponding to areas around the polymer remains was noted in the animals treated with copolymer loaded with the growth hormone. Copolymer is biodegradable in vivo and may be used as a vehicle for the slow release of active substances. The presence of the hormone at the site of skin injury induces cell proliferation and enhances the repair process.

Tissue integration and biomechanical behaviour of contaminated experimental polypropylene and expanded polytetrafluoroethylene implants.

BACKGROUND: Infection is one of the most devastating complications following implantation of a prosthetic material. The aim of this study was to compare the behaviour of two biomaterials contaminated with Staphylococcus aureus or Staphylococcus epidermidis, used to repair abdominal wall defects. METHODS: Defects (7 x 5 cm) were created in the anterior abdominal wall of 60 white New Zealand rabbits and repaired using polypropylene or expanded polytetrafluoroethylene (ePTFE) prostheses. The site of repair had been previously inoculated with 10(6) colony-forming units/ml S. aureus or S. epidermidis. Seven and 30 days after implantation, prosthetic specimens were examined by light and scanning electron microscopy, and immunohistochemical and biomechanical analysis. RESULTS: No significant differences with respect to controls were observed in the S. epidermidis groups. Two animals inoculated with S. aureus died. S. aureus induced the appearance of denuded areas exposing the filaments in the polypropylene prostheses, whereas the ePTFE prostheses showed zones of erosion, disorganized tissue, haemorrhage and necrosis. The biomechanical strength of the contaminated implants was unaltered. CONCLUSION: Integration within host tissue was affected in the setting of S. aureus infection but the tensile strength of contaminated prostheses was not significantly reduced.

Long-term behaviour of cryopreserved arterial grafts versus prosthetic micrografts.

INTRODUCTION: When a patient has no suitable vessels for use as grafts in bypass or reconstruction procedures, two of the options available are the use of a cryopreserved vessel or an expanded polytetrafluoroethylene (ePTFE) prosthesis. This study was designed to compare the long-term behaviour of these vascular substitutes. MATERIAL AND METHODS: We established three study groups by grafting the following vessel substitutes to the iliac artery in Spraque-Dawley rats: arterial autografts (GI, n=12), cryopreserved syngenic arterial grafts (cryoisografts) (GII, n=12), and ePTFE micrografts (GIII, n=12). The animals were sacrificed 180 days after surgery, at which time the graft specimens were morphologically evaluated by light and electron microscopy, immunolabelling (ED1/alpha-actin) and morphometric analysis of the neointima. RESULTS: At the time of sacrifice, graft patency was 100% for the autografts and cryoisografts, while 10% of the ePTFE micrografts showed fully-occlusive thrombosis. Intimal hyperplasia was observed in grafts in GI and GII; the neointima being thinner in the cryoisografts (54.36 +/- 2.26 microm) than the autografts (161.30 +/- 3.91 microm). The endothelium formed over the prosthetic micrografts was unstable, with areas of subendothelial thickening (9.37 +/- 3.18 microm). Cell loss and medial layer degeneration were observed in both GI and GII specimens, while the GIII grafts were colonised by cells on their luminal surface. CONCLUSIONS: All three grafts show good long-term tolerance when used in an arterial setting. Following long-term implant, autografts and cryoisografts show similar alterations that give rise to the complete loss of the muscle component of the tunica media along with the formation of a stable neointima. This new layer takes on the role of the tunica media.

Isolation of intact aortic valve scaffolds for heart-valve bioprostheses: extracellular matrix structure, prevention from calcification, and cell repopulation features.

Extracellular matrix (ECM) scaffolds isolated from valvulated conduits can be useful in developing durable bioprostheses by tissue engineering provided that anatomical shape, architecture, and mechanical properties are preserved. As evidenced by SEM, intact scaffolds were derived from porcine aortic valves by the combined use of Triton X-100 and cholate (TRI-COL) or N-cetylpyridinium (CPC) and subsequent nucleic acid removal by nuclease. Both treatments were effective in removing most cells and all the cytomembranes, with preservation of (1) endothelium basal membranes, (2) ECM texture, including the D-periodical interaction of small proteoglycans with normally D-banded collagen fibrils, and (3) mechanical properties of the treated valves. Ultrastructural features agreed with DNA, hexosamine, and uronic acid biochemical estimations. Calcification potential, assessed by a 6-week rat subdermal model, was significantly reduced by TRI-COL/nuclease treatment. This was not true for CPC only, despite better proteoglycan preservation, suggesting that nucleic acids also are involved in calcification onset. Human fibroblasts, used to repopulate TRI-COL samples, formed mono- or multilayers on surfaces, and groups of cells also were scattered within the valve leaflet framework. A biocompatible scaffolds of this kind holds promise for production of durable valve bioprostheses that will be able to undergo probable turnover and/or remodeling by repopulating recipient cells.

A novel controlled drug-delivery system for growth hormone applied to healing skin wounds in diabetic rats.

Controlled release systems for drugs, hormones and growth factors can be particularly useful in tissue repair processes. These systems act as a biodegradable support containing the substance to be delivered, allowing their gradual release. In the past years, the local application of growth factors has acquired special relevance as a therapeutic option for use in subjects who show deficient tissue scarring, the hormone dose being the limiting factor for its success. In this study, the in vitro biocompatibility of a copolymer formed by vinylpyrrolidone and 2-hydroxyethyl methacrylate, used as an administration vehicle for hGH, was evaluated. The system was then tested in vivo in terms of its capacity for healing incisional wounds in healthy and diabetic rats. For the in vitro studies, polymer and hormone degradation rates were determined, and polymer biocompatibility was evaluated in fibroblast cultures. In the in vivo experiments, an incision was made in the back of the animals, and polymers discs with/without hGH, were introduced in the aperture. Morphological, immunohistochemical and morphometric evaluations were performed on wound tissue specimens 3-10 days after surgery. In vitro, the polymer was found to be biodegradable and showed no toxic effects on fibroblasts, the hormone being slowly released to the culture medium. In untreated diabetic rats, a delayed skin scarring and cell response were observed, compared to that noted in healthy animals. Skin closure, keratinisation and fibrosis occurred earlier in the presence of the polymer-hGH system. The use of this co-polymer as an administration vehicle for hGH improves the wound scarring process in the pathological setting of diabetes.

Modulatory role of IL10 in endothelial cell damage and platelet adhesion.

This study explores the possibility of a regulatory role for cytokine IL-10 in platelet aggregation as an active vascular repair mechanism. Endothelial cells from human umbilical cord vein were cultured in the presence of different IL-10 concentrations (0-100 ng/ml). Platelet-rich plasma was then added to these cultures and allowed to act for 30 minutes. To rule out blood plasma involvement, washed platelets were also incubated with IL-10 (0-100 ng/ml). Changes in endothelial cell morphology were observed depending on the IL-10 concentration used; apoptotic cells appearing at the highest IL-10 concentration. Greatest platelet adhesion was noted at the highest IL-10 concentration. It was concluded that, in this in vitro model, low IL-10 levels do not affect cell viability or the pattern of platelet adhesion, but at high doses, this cytokine induces cell death and enhances platelet deposition.

In vitro mesothelialization of prosthetic materials designed for the repair of abdominal wall defects.

The aim of this study was to evaluate the in vitro response of mesothelial cells (MC) in terms of their ability to cover different biomaterials. MC were harvested from human omentum. The MC from the first passage were seeded onto different biomaterials from 10 min to 24 h: PL-PU99 (polypropylene-polyurethane); DM (ePTFE); PL (polypropylene); and PL + Col (polypropylene-collagen). The prosthetic surface covered was examined by microscopy and quantified. PL-PU99: The MC were adhered to the biomaterial 10 min post-incubation. At 4 h, the 53.12+/-7.86% of the prosthesis were coated with polygonal cells. At 12 h, 96.32+/-11.32% of the biomaterial was coated. DM: between 30 min to 8 h, the MC cells form small, round colonies. At 12 h, polygonal and fusiform secretory cells were observed (68.94+/-5.78%). 93.54+/-11.49% of surface was coated after 24 h. PL: only isolated cells were observed on the prosthesis. PL + Col: MC form a monolayer over prosthetic surface after 18 h (90.21+/-9.76). We conclude: (a) MC formed a stable monolayer over all the biomaterials tested with the exception of the PL due to its porosity. (b) The PL-PU99 showed the greatest potential for in vitro mesothelialization compared to the PL-Col and DM prostheses.

Estudio experimental de homoinjertos criopreservados: modificaciones miointimales

Objetivo. Evaluar el desarrollo de hiperplasia intimal, muerte celular y respuesta macrofágica, en injertos arteriales criopreservados tras su implante en animales de experimentación. Material y métodos. Arterias ilíacas de rata Spraque-Dawley fueron criopreservadas de manera automatizada, en un congelador biológico, con un rango de disminución de temperatura de 1ºC/min. Fueron almacenadas a 145ºC en vapores de N2 líquido durante 30 días y sometidas a descongelación lenta y computerizada. Los injertos singénicos, se efectuaron a nivel de la arteria ilíaca común. Se establecieron los siguientes grupos de estudio: GI: arterias criopreservadas no implantadas, GII: microinjertos frescos implantados y GIII: arterias criopreservadas e implantadas. Manteniendo como grupo control (GC), arterias ilíacas frescas. Los animales fueron sacrificados a los 14, 30 y 90 días post-injerto. Se valora la hiperplasia intimal sobre los injertos mediante morfometría, se realizan técnicas de TUNEL y respuesta macrofágica. Resultados. La permeabilidad en el momento del sacrificio fue del 100 per cent para homoinjertos fresco y 66.6 per cent para los implantes criopreservados. El propio proceso de criopreservación (GI) indujo daño en la pared arterial sobre todo a nivel endotelial mostrando áreas de denudación de diferente extensión y una buena celularidad de la capa media. La hiperplasia intimal sobre el injerto a los 14 días de implante (GII), estaba visiblemente retrasada hasta los 30 días cuando el injerto era criopreservado (GIII). Las arterias de este último grupo, mostraban un adelgazamiento general de la pared arterial y degeneración con pérdida de celularidad a nivel de la capa media del injerto. La presencia macrofágica fue similar en los dos grupos implantados, limitándose a zonas de anastomosis e hiperplasia. Los estudios de daño celular mostraron mayor número de células marcadas en el GIII a los 90 días post-implante. Conclusiones. La respuesta hiperplásica se modifica cuando el injerto ha sido previamente criopreservado, el retraso en su formación y una mayor tasa de muerte celular a largo plazo, sin alteraciones en la respuesta macrofágica, parecen ser indicativos de una clara tendencia a la degeneración del mismo a largo plazo (AU)

Caracterización y biocompatibilidad de las prótesis vasculares de poliuretano estabilizado con polidimetilsiloxano / Characterization and biocompatibility of vascular prostheses made of polyurethane stabilized with polydimethylsiloxane

Introducción. El desarrollo de nuevos biomateriales ha desembocado en la aparición de nuevas prótesis vasculares que mejoren el comportamiento de injertos protésicos de pequeño calibre. Objetivo. El objetivo del presente trabajo es el estudio del comportamiento biológico de prótesis vasculares de poliuretano. Material y métodos. Prótesis: poliuretano-polidimetilsiloxano (PU-PDMS). Caracterización: fragmentos de PU-PDMS se procesaron para su estudio en microscopia óptica y electrónica de barrido. Se determinó la carga eléctrica de la superficie interna mediante análisis espectral. Biocompatibilidad: fragmentos (1 cm2) de PU-PDMS se implantaron en el músculo dorsal de conejos Nueva Zelanda (n= 18) durante 3 y 8 meses. Realizamos estudios morfológicos, inmunohistoquímicos (antiactina) y de reacción de cuerpo extraño (RAM11). Siembra: fragmentos de 1 cm2 se sembraron con células endoteliales de vena umbilical humana. Tiempos de estudio: 24, 48, 72 horas y 7 días. Resultados. La composición es fibrilar, con presencia de numerosos poros. Existencia de cargas negativas en la superficie interna del biomaterial. A los tres meses, la prótesis se embebe en tejido neoformado muy vascularizado y rico en células blancas y células de reacción a cuerpo extraño. A los 8 meses se puede observar la total integración del biomaterial, que aparece rodeado de colágeno y muy vascularizado. A las 24 horas de la siembra observamos una superficie endotelizada, que deja al descubierto grandes poros que se tapizan en los estadios posteriores. Conclusiones. Las prótesis PUPDMS presentan características adecuadas para utilizarse como sustitutos vasculares, gracias a su estructura, ausencia de rechazo y buena integración a corto y medio plazo. (AU)

Polymer controlled drug delivery system for growth hormone.

The use of biomaterials as vehicles for pharmacological agents, hormones, and growth factors is at times the best treatment for controlled local administration. Our study was designed to evaluate the in vitro biocompatibility and potential clinical use of a new polymer, hydroxyethyl methacrylate-vinyl pirrolidone. Human fibroblasts were incubated in the presence of the polymer and/or growth hormone, and evaluation was made of both the rate of polymer and hormone degradation and the proliferative effect on the fibroblast population. Results indicate that this polymer is biodegradable and lacks toxicity toward these cells. The hormone was slowly released, as suggested by enhanced cell proliferation.

Diseño y ensayo biológico de una nueva prótesis composite (PL-PU99) destinada a la reparación de defectos de la pared abdominal / Design and biological trial of a new composite prosthesis (PL-PU99) for the repair of abdominal wall defects

Introducción. Las prótesis macroporosas, tipo polipropileno (PL), empleadas para la reparación de defectos en la pared abdominal, tienen en algunas ocasiones que ser implantadas en contacto con el peritoneo visceral. La interfase prótesis/peritoneo visceral puede generar problemas en cuanto a formación adherencial con posibilidad de formación de fístulas. El objetivo del presente trabajo ha sido realizar un estudio sobre el comportamiento en esta interfase de una nueva prótesis diseñada en forma de composite (PL-PU99) por nuestro grupo de investigación. Material y métodos. Se han empleado 30 animales (conejo blanco Nueva Zelanda) de un peso aproximado entre 2.000 y 2.500 g. Se crearon defectos de 7 × 5 cm en la pared anterior del abdomen que comprendían todos los planos (aponeurótico, muscular y peritoneo parietal), siendo reparados los mismos con prótesis de PL y PL-PU99. La piel que quedó cubriendo la prótesis fue cerrada con una sutura de polipropileno 3/0. La prótesis PL-PU99 es un composite formado por tres componentes: una prótesis de PL de un poro de 1 mm y una lámina de poliuretano (colocada en contacto con el peritoneo visceral), unidas ambas por un pegamento acrílico. Se establecieron dos grupos de estudio: grupo I (n = 15) o control, implantes de PL, y grupo II (n = 15), implantes de PL-PU99. Los animales fueron sacrificados a los 14, 30 y 90 días de la intervención quirúrgica. Se efectuaron estudios a microscopia óptica, electrónica de barrido (SEM) y transmisión (MET), inmunohistoquímica y morfometría del neoperitoneo. Asimismo, se cuantificaron las adherencias en la interfase prótesis/peritoneo visceral. El estudio biomecánico se realizó con un tensiómetro Instron (TT-DM-1118). El análisis estadístico se efectuó empleando los test de la t de Student-Newman-Keuls y la U de Mann-Whitney. Resultados. No hubo mortalidad en los animales intervenidos ni presencia de infección o rechazo de los implantes. Las adherencias fueron firmes en los implantes de PL y prácticamente inexistentes en los de PL-PU99. La superficie cubierta por adherencias fue de 7,18 ñ 1,11 y 0,11 ñ 0,02 cm2, respectivamente, para los grupos I y II, existiendo diferencias significativas entre ambos grupos (p < 0,01). El neoperitoneo formado en el grupo I presentó una disposición anárquica y desordenada, de textura rugosa. En algunas ocasiones pudieron apreciarse zonas de hemorragia y necrosis que se correspondían con las zonas en las que se habían producido adherencias. Por el contrario, en el grupo II fue homogéneo, conformado por un tejido conectivo ordenado y vascularizado, todo ello tapizado por un mesotelio. El análisis morfométrico del neoperitoneo fue significativamente mayor (p < 0,05) en el PL-PU99 (474,86 ñ 49,73) que en el PL (256 ñ 21,68).Los resultados del estudio inmunohistoquímico demostraron características similares en los dos tipos de implantes. La evaluación de la resistencia biomecánica no evidenció diferencias significativas entre las dos prótesis en los distintos tiempos de estudio. Conclusiones. a) la prótesis PL-PU99 tiene un comportamiento óptimo en cuanto a formación adherencial, en la interfase prótesis/peritoneo visceral; b) el neoperitoneo formado con esta prótesis sustituye casi física y funcionalmente al peritoneo normal, y c) la resistencia biomecánica obtenida no presenta diferencias entre el grupo control y el grupo objeto de estudio (AU)