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Biomedical subjects

S H Hyon

Publications and source records attributed to S H Hyon.

At least 19 recordsLinked to original sources

Trypanosoma cruzi trans-sialidase inhibits human lymphocyte proliferation by nonapoptotic mechanisms: implications in pathogenesis and transplant immunology.

Chiefly an intracellular parasite, Trypanosoma cruzi has a transient blood-borne stage (trypomastigote), the acute phase of Chagas' disease, during which surface trans-sialidase is expressed and shed by the parasite. It's immunosuppressive through the induction of apoptosis. Herein, we investigated the role of trans-sialidase as an immune modulator of allo- and xenoreactions. Trans-sialidase strongly inhibited human lymphocyte proliferation; a role for the interleukin-2 receptor CD25 was suggested by flow cytometry. These results may have implications both for the pathogenesis of Chagas' disease and for transplantation immunology.

Animals↗

Protective effects of green tea polyphenol against reactive oxygen species-induced oxidative stress in cultured rat calvarial osteoblast.

The injurious effects of reactive oxygen species on osteoblasts and the potential protective role played by green tea polyphenols (GtPP) were investigated using primarily cultured rat calvarial osteoblasts. Oxidative stress was induced in cultured osteoblasts, either by adding 100 mmol/L H2O2 or by the action of 40 U/L xanthine oxidase (XO) in the presence of xanthine (250 micromol/L). After incubation, the cellular viability, function and morphology were evaluated. Both treatments produced a significant reduction in osteoblast viability, as assessed by a two-colored fluorescence staining method combined with flow cytometric analysis and MTT assay. A significant reduction in the alkaline phosphatase activity was observed after H2O2 addition, whereas XO did not have the same effect. On the microscopic observations, the morphological changes and intracellular ultrastructural damages were remarkably induced by both treatments. The H2O2-induced alterations were prevented by pre-incubating the osteoblasts with 200 microg/ml GtPP for 1 h. When the oxidative stress was induced by XO, the cellular viability and morphology was also maintained at the same polyphenol concentration. These results demonstrate that GtPP can act as a biological antioxidant in a cell culture experimental model and protect cells from oxidative stress-induced toxicity.

Alkaline Phosphatase↗

Partial hemiarthroplasty for the treatment of osteonecrosis of the femoral head. An experimental study in the dog.

The use of a composite osteochondral device for simulating partial hemiarthroplasty was examined. The device was composed of a polyvinyl alcohol hydrogel and a titanium fibre mesh, acting as artificial cartilage and as porous artificial bone, respectively. The titanium fibre mesh was designed to act as an interface material, allowing firm attachment to both the polyvinyl alcohol gel (through injection moulding) and the femoral joint surface (through bony ingrowth). We implanted 22 of these devices into canine femoral heads. Histological findings from the acetabular cartilage and synovial membrane, as well as the attachment of the prosthesis to bone, were examined up until one year after operation. No marked pathological changes were found and firm attachment of the device to the underlying bone was confirmed. The main potential application for this device is for partial surface replacement of the femoral head after osteonecrosis. Other applications could include articular resurfacing and the replacement of intervertebral discs.

Acetabulum↗

Long-term preservation of rat pancreatic islets under physiological conditions.

In this study, we found that islet cells treated with polyphenol could be preserved for over 2 months under physiological conditions retaining their original function and maintaining their spherical shapes without any insulin secretion. When islets were treated at higher concentration than 250 microg ml(-1), these islets could retain their compact spherical shape over 65 days whereas non-treated islets were scattered ease to break within 2 weeks. The secretional capacity from treated islets in the initial stage is also lower than untreated islets. However, in the case of untreated islets, insulin release rapidly lowered with the progress in the culture time and secretion completely disappeared after 9 days. On the contrary, islets treated with polyphenol (250 microg ml(-1)) in RPMI culture medium showed significant enhancement of insulin secretion on 40th day. The secretional capacity of islets was greatly dependent on the treating concentration. Polyphenol treatment may be a useful method for preservation of mammalian islet cells. By changing the concentration of polyphenol, it is possible to control the preservation duration and insulin secretion of islets.

Animals↗

Morphologic study and syntheses of type I collagen and fibronectin of human periodontal ligament cells cultured on poly(ethylene-co-vinyl alcohol) (EVA) with collagen immobilization.

The purpose of this study was to regenerate the human periodontal ligament (PDL) around dental implants by using a hybrid structure of cells with materials, such as PDL cells + collagen + poly(ethylene-co-vinyl alcohol) (EVA) on titanium implant. Human PDL cells were cultured on the EVA surface coated with type I collagen and the cell adhesion and extension were investigated. Furthermore, collagen type I and fibronectin syntheses were analyzed. The serum free culture was also tried, to investigate the role of collagen in detail. The results showed that: 1. Satisfactory adhesion, extension, and proliferation of the PDL cells on the EVA films coated with collagen were observed, but were not good without collagen. 2. Immunostaining of cultured PDL cells revealed the syntheses of type I collagen, when cultured on the EVA coated with collagen or conventional culture dish, though fibronectin synthesis was observed even in the EVA without collagen. 3. Only PDL cells on the EVA coated with collagen proliferated well in the absence of serum. These results indicate that our novel implant material (EVA coated with collagen) provides a possibility of PDL regeneration on dental implants.

Cell Culture Techniques↗

Long-term implantation test and tumorigenicity of polyvinyl alcohol hydrogel plates.

Two types of flat plates made from a polyvinyl alcohol (PVA) hydrogel with a water content of 80 and 20 (PVA-H80, PVA-H20), 20 x 10 x 1 mm in size, were subcutaneously implanted into each of 50 young, male Wistar rats. As a control, a sham operation was done on another set of 50 rats (Sham Op group). The shape and transparency of the PVA hydrogel were unchanged for up to 24 months. Tumors arose in 14 rats from the PVA-H80 group. In the PVA-H20 group, tumors appeared in 15 rats. The average tumor latency was 598 +/- 109 days in the PVA-H80 and 637 +/- 94 days in the PVA-H20. There was no difference in tumor incidence between the PVA-H20 and PVA-H80 groups (p < 0.05). In the Sham Op group, no malignant tumors appeared. Histopathologically, the tumors induced by hydrogel plates were malignant tumors resembling fibrosarcoma or malignant fibrous histiocytoma. This indicates that PVA hydrogel implants also induce solid state carcinogenesis at a similarly high rate to medical grade hydrophobic material reported in a previous study.

Absorbable Implants↗

Culture of porcine hepatocytes: the dogma of exogenous matrix revisited.

The use of exogenous matrices has been described as an essential component in securing the viability and functionality of hepatocytes in vitro whether cultured for extracorporeal devices or cell transplantation. Here we report on the in vitro culture of porcine hepatocytes in polystyrene tissue-culture flasks without exogenous matrices showing adequate attachment and viability. Cell proliferation was evidenced by uptake of 5-bromo-2'-deoxyuridine, with peaks at Days 2 (19.7 +/- 8.5%), 15 (20.8 +/- 3.3%), and 35 (21.4 +/- 0.3%). Detoxification capacity was assessed by determination of monoethylglycinexylidide, a product of lidocaine metabolism (highest value 156.5 +/- 10.1 ng/ml at Day 4), and by diazepam clearance (maximum clearance 66.2% at Day 6). Diazepam metabolite levels were highest at Day 4 both for temazepam and oxazepam (6.5 +/- 0.1 and 0.10 +/- 0.01, respectively). These results suggest that the need for an exogenous matrix to achieve sustained proliferative activity and differentiated hepatocyte function should not necessarily be considered a sine qua non condition.

Animals↗

Tissue-engineered pancreatic islets: culturing rat islets in the chitosan sponge.

Subcutaneous islet transplantation has become an attractive modality. With development of tissue-engineering techniques, it is possible to rectify the disadvantage of poor blood supply in the subcutaneous site by reconstruction of the capillary network. According to reports, the Chitosan sponge (CS) could be used for reconstruction of in vitro capillary-like network and could be used in artificial skin equivalent. In this study, we cultured the islets in CS for future application. CSs, having 200-500 microm pore size, were prepared by freeze-drying method. Rat islets were isolated from the pancreas of Lewis rats (10 weeks old, 280-300 g, male) by collagenase digestion followed by discontinuous dextran gradient centrifugation method. Each 20 islets were seeded equally into the CSs and were cultured for 62 days with various culture media such as RPMI-1640, Dulbecco's modified Eagle's medium (DMEM), and Eagle's MEM. They contained 10% fetal bovine serum (FBS) and 5 ml/L antibiotic-antimycotic mixed stock solution in the culture dishes. Insulin concentration both inside and outside of the islet-seeded CS was measured during culture. Changes in the morphology of islets were also observed in this study. Freshly isolated islets had a loose appearance with an irregular border, and most were seen as a single islet. Occasionally a cluster, consisting of 2-4 islets ranging mainly from 150 to 250 microm in diameter, was observed. Islets cultured in the CSs in different culture media retained initial morphology, which had well-delineated smooth borders for at least 53 days. The insulin release behavior of islets cultured in the CS showed constant secretory capacities for 49 days. After that they exhibited a rapid and definitive decline from the initial insulin release. Until this stage, insulin concentration in the CS was well maintained. The properties were dependent on culture medium used and insulin diffusion released from islets. This experiment is a new study model for establishment of islet culture in a three-dimensional matrix. Also extension of this observation will provide new insights for islet transplantation at the subcutaneous site by a tissue-engineering approach.

Animals↗

Type I atelocollagen grafting onto ozone-treated polyurethane films: cell attachment, proliferation, and collagen synthesis.

An approach is presented for the graft copolymerization of type I atelocollagen onto the surface of polyurethane (PU) films treated with ozone. Through inducing oxidization to modify PU surface by ozone, peroxide groups are easily generated on the surface. Those peroxides are broken by redox-polymerization, and provide active species which initiate graft polymerization by reacting with amines in the collagen molecules. The ozone oxidation time and voltage could readily control the amount of peroxide production. The surface density of generated peroxides on PU surface was determined by iodide method. The maximum concentration of peroxide was about 10.20 x 10(-8)mol/cm(2) when ozone oxidation was performed at 60 V for 30 min. After the reaction of PU by ozone oxidation, type I atelocollagen was graft-copolymerized onto the PU film. All the physical measurements on the collagen-grafted surface indicated that the PU surface was effectively covered with type I atelocollagen. The interaction of the collagen-grafted PU surface with fibroblasts could be greatly enhanced by the surface graft polymerization with type I atelocollagen. Attachment and proliferation of fibroblasts on the grafted type I atelocollagen were significantly enhanced, and it is assumed that the atelocollagen matrix supported the initial attachment and growth of cells. In the early stage of proliferation, collagen synthesis in fibroblasts was not activated and remained at a relatively low level due to the grafted type I atelocollagen, increasing only with fibroblast differentiation.

Animals↗

Biodegradable scleral implant for controlled intraocular delivery of betamethasone phosphate.

We evaluated nail-like, biodegradable scleral implants as a controlled intraocular delivery system of betamethasone phosphate (BP) for the treatment of chronic uveitis using pigmented rabbits. The scleral implants, which contained 10% and 25% of BP (weight 8.5 mg; length 5 mm), were made of poly (D,L-lactide-co-glycolide) (PLGA). In vivo release and retinal toxicity after implantation were also studied in pigmented rabbits. The in vitro release studies demonstrated the 10% and 25% BP-loaded scleral implants released BP in a biphasic release pattern for at least 1 month. The BP concentrations in the vitreous and the retina/choroid after application of scleral implants in pigmented rabbit eyes stayed within the concentration range capable of suppressing inflammatory responses for over 1 month. The BP concentration was greater in the retina/choroid than in the vitreous throughout the study. No substantial toxic reactions in the retina were observed by electroretinography. Our findings suggest that the BP-loaded scleral implant may be a promising device for treatment of chronic uveitis.

Animals↗

Surface modification of poly(ethylene-co-vinyl alcohol) (EVA). Part I. Introduction of carboxyl groups and immobilization of collagen.

To enhance the surface biocompatibility of poly(ethylene-co-vinyl alcohol) (EVA) and high-density polyethylene (HDPE), carboxyl groups were introduced by ozone exposure. Type I collagen was immobilized onto the surface through polyion complexing. The carboxyl groups on the EVA were characterized by electron spectroscopy for chemical analysis and neutralization. The amounts of the carboxylic group and collagen increased with increases in time and temperature of exposure. Water-soluble fragments were produced by ozone exposure to EVA, and they acted as collagen crosslinkers. The differences in charge distribution of carboxyl groups affected the amount of collagen immobilization. Graft polymerization of acrylic acid was also carried out onto EVA and HDPE surfaces. The amount of collagen immobilized by graft polymerization was much higher than that by ozone exposure despite the introduction of almost the same amounts of carboxylic groups. It was suggested that the negative charge distribution influences the amount of collagen immobilized onto films.

Biocompatible Materials↗

Development of artificial articular cartilage.

Attempts have been made to develop an artificial articular cartilage on the basis of a new viewpoint of joint biomechanics in which the lubrication and load-bearing mechanisms of natural and artificial joints are compared. Polyvinyl alcohol hydrogel (PVA-H), 'a rubber-like gel', was investigated as an artificial articular cartilage and the mechanical properties of this gel were improved through a new synthetic process. In this article the biocompatibility and various mechanical properties of the new improved PVA-H is reported from the perspective of its usefulness as an artificial articular cartilage. As regards lubrication, the changes in thickness and fluid pressure of the gap formed between a glass plate and the specimen under loading were measured and it was found that PVA-H had a thicker fluid film under higher pressures than polyethylene (PE) did. The momentary stress transmitted through the specimen revealed that PVA-H had a lower peak stress and a longer duration of sustained stress than PE, suggesting a better damping effect. The wear factor of PVA-H was approximately five times that of PE. Histological studies of the articular cartilage and synovial membranes around PVA-H implanted for 8-52 weeks showed neither inflammation nor degenerative changes. The artificial articular cartilage made from PVA-H could be attached to the underlying bone using a composite osteochondral device made from titanium fibre mesh. In the second phase of this work, the damage to the tibial articular surface after replacement of the femoral surface in dogs was studied. Pairs of implants made of alumina, titanium or PVA-H on titanium fibre mesh were inserted into the femoral condyles. The two hard materials caused marked pathological changes in the articular cartilage and menisci, but the hydrogel composite replacement caused minimal damage. The composite osteochondral device became rapidly attached to host bone by ingrowth into the supporting mesh. The clinical implications of the possible use of this material in articular resurfacing and joint replacement are discussed.

Aluminum Oxide↗

Biodegradable poly (lactic acid) microspheres for drug delivery systems.

In connection with aim of maximizing the bio-availability of conventional drugs with minimum side-effects, new drug delivery systems (DDS) continue to attracted much attention. The controlled or sustained release of drugs represents one such approach, and in this regard report upon a study of DDS using biodegradable polymers which include poly (lactic acid) (PLA), poly (glycolic acid), and their copolymers (PLGA). Much attention is being paid to the controlled release of bio-active agents from microcapsules and microspheres made of biodegradable polymers, such as lactic acid homopolymers, as well as copolymers of glycolic acid. (11-21) Microcapsules or microspheres are injectable and able to provide pre-programmed durations of action, offering several advantages over the conventional dosage forms. This article reviews the results of a work program conducted in collaboration with a medical doctor upon DDS using biodegradable microspheres, such as PLA and PLGA.

Animals↗

[Kidney and pancreas transplantation: initial experience at a single transplant center in Argentina].

After more than 10,000 cases reported all over the world until 1998, simultaneous kidney and pancreas transplantation has become a safe clinical practice, and it may probably represent the best treatment available for diabetic patients in end-stage renal disease. Here we present our results after 12 cadaveric pancreas transplants (8 whole organ, and 4 islet transplants), performed on insulin-dependent diabetic patients. Eleven of these patients received a kidney simultaneously, and one of them required a kidney retransplantation. All vascularised pancreatic grafts were positioned intraperitoneally, anastomosed to the iliac vessels, and bladder drained. One year patient, whole pancreas, and kidney survival rates were 86%, 86% and 71%, respectively. All of these patients remain insulin and dialysis-free, the longest for 37 months. Islets for transplantation were obtained from single cadaveric donors. Fresh, unpurified cells were transplanted intraperitoneally by laparoscopy (equivalent islet yields: 3 x 10(5), 4 x 10(5), 1 x 10(6) and 5 x 10(5)). None of the islet recipients resulted insulin-independent but they all reduced daily requirements in about 40%, with better metabolic control (mean HbA1c pretransplant 9.4 +/- 1.8, vs 7.9 +/- 1.6 posttransplant). One kidney graft was lost due to venous thrombosis. Simultaneous kidney and pancreas transplantation offers the diabetic patient in end-stage renal disease a chance of independence both from dialysis and exogenous insulin. Whole pancreas transplantation has better functional outcome than islet transplantation. Nevertheless, for those diabetic patients who do not meet the criteria to receive a vascularised graft, pancreatic cells may still improve carbohydrate metabolism with minor surgical risk.

Adult↗

Biodegradation and tumorigenicity of implanted plates made from a copolymer of epsilon-caprolactone and L-lactide in rat.

Flat plates made from a copolymer of epsilon-caprolactone and L-lactide (P-CL-LA) [50:50 (w/w), molecular weight 1.62 x 10(5); 20 x 10 x 1 mm size] were subcutaneously implanted into 50 young, male Wistar rats (P-CL-LA group). After 24 months the plates had become a mass of small pieces, which were concentrated in an area of 3 x 2 x 1 mm. For comparison, 50 rats were implanted with medical-grade polyethylene plates (PE group) while another set of 50 rats was subjected to the same operation but without an implant (Sham Op group). Tumors arose in 25 rats from the P-CL-LA group: 24 were malignant mesenchymal tumors at the implant sites. In the PE group, tumors appeared in 16 rats (14 at the implant sites and two ectopically). The average tumor latency was 578+/-84 days in the P-CL-LA group and 452+/-102 days in the PE group. There was no difference in tumor incidence between the P-CL-LA and PE groups (p < 0.05). In the Sham Op group, two malignant tumors appeared over 2 years. Pathologically, these induced tumors arose from the inflammatory cells surrounding the degrading fragments of P-CL-LA within the tissue capsule. This indicates that relatively slowly degrading material can induce malignant tumors at a similarly high rate to nonabsorbable medical grade PE, at least in this animal model.

Abdominal Neoplasms↗

Occlusive effects of lactic acid-glycolic acid copolymer membrane on gingival fibroblasts in vitro.

The cell occlusive effects on human gingival fibroblasts of degradable lactic acid-glycolic acid copolymer membranes (noncoated membranes) and membranes coated with a sucrose ester of fatty acid (coated membranes) were studied and compared with those of expanded polytetrafluoroethylene (e-PTFE) membranes. The membranes were immersed in a culture medium periodically for 21 days and interposed into a chemotaxis chamber, and the fibroblasts then were cultured in the chamber for another 7 days. The passage rate of cells through the membranes was calculated and the change in surface structure of each membrane after immersion for 28 days was observed by an environmental scanning electron microscope. The passage rate of coated membranes (3.4+/-2.2%) was significantly lower than that of noncoated (25.7+/-5.1%) at the 28th day whereas the passage rate of e-PTFE membranes was 0.8-1.5%. Many pores were observed on the noncoated membranes before immersion while the coating material covered most of the pores on the coated membranes. The average pore size of the noncoated membranes was larger than that of the coated membranes at day 28. The structure of the e-PTFE membranes underwent no change. The passage rate of the coated membranes was not different from the e-PTFE membranes, suggesting an effect that might be useful for a guided tissue regeneration procedure.

Fibroblasts↗