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The use of silicone/polyurethane graft polymers as a means of eliminating surface cracking of polyurethane prostheses.

The long-term biodegradation of various polyurethanes with and without surface modifications was evaluated by implanting small porous filamentous patches of these materials subcutaneously in the backs of dogs for one month. Data were compared to those obtained with spun polyurethane vascular grafts of similar materials implanted in the aorto-iliac position in dogs. The extremely high surface area of approximately 7 m2/cm3 of these porous filamentous patches provided numerous sites for surface cracking and the very fine filaments (10 microns in diameter) provided an easily identifiable structure to study the cracking phenomenon. Results from numerous one month implants clearly demonstrated that the subcutaneous implant model effectively reproduced the biodegradation behavior observed in vascular graft implants. The degradation was most pronounced in the softer Shore 80A polyurethanes and less pronounced in the harder 55D and 75D polyurethanes. The degradation could not simply be stopped by stress annealing the polyurethane and the degradation did not require the presence of metallic ions. Antioxidants, surface adsorbed albumin, poly(2-hydroxyethyl-methacrylate) grafting, silicone copolymerization, tetrafluoroethylene plasma discharge and the addition of urea linkages to the polymer were also shown to be ineffective in stopping the biodegradation process. In contrast, covalent bonding or grafting of silicone polymer to the surface of the urethane successfully inhibited the biodegradation process.

Animals

Covalent immobilization of proteins on to the surface of poly(vinyl alcohol) hydrogel.

Covalent immobilization of cell-adhesive proteins such as collagen and fibronectin on to the surface of poly(vinyl alcohol) hydrogel was investigated by using diisocyanates, polyisocyanates, and cyanogen bromide. It was found that 0.5 and 12 micrograms/cm2 of collagen were immobilized on to the surface by using hexamethylene diisocyanate and cyanogen bromide, respectively. The big difference in the graft amount between the two methods was ascribed to the different reaction media employed for the surface modifications; toluene for the reaction with hexamethylene diisocyanate and water for the reaction with cyanogen bromide.

Collagen

Hydrophilic albumin microspheres.

A method has been developed for preparing unique hydrophilic HSA/MS. Important aspects of this synthesis include addition of the cross-linking agent (glutaraldehyde) in the organic phase and use of concentrated polymer solutions as dispersion media. The polymer solutions afford excellent steric stabilization of aqueous albumin microdispersions for microsphere synthesis. Steric stabilization of dispersions by polymer solutions was shown to be a function of polymer concentration and molecular weight. The HSA/MS prepared by this method are hydrophilic and easily dispersed in a variety of aqueous media without surfactants. Chemical modifications are easily accomplished using available reactive aldehyde groups remaining after cross-linking. Although hydrophilicity of the microspheres is advantageous for many drug delivery applications, in some instances (such as the use of MS in adjuvant immunotherapy or vaccine preparations) some hydrophobicity may be desirable. For this purpose, surface modifications to produce controlled hydrophobicity is easily achieved by covalent coupling with appropriate reagents (e.g., fatty amines). Adriamycin was bound to HSA/MS by both physical association (to 18 wt%) and covalent binding (also to 18 wt%). In vitro release of drug was measured for the MS using a dynamic flow method. Two distinct release mechanisms could be achieved depending on the type of drug bonding used: slow by hydrolytic degradation of covalent bonds and fast by release of physically adsorbed drug. This new and versatile synthesis of hydrophilic HSA/MS opens up many new opportunities for producing chemically modified MS containing high concentrations of therapeutic agents. Use for immunodiagnostic and adjuvant compositions is also suggested.(ABSTRACT TRUNCATED AT 250 WORDS)

Capsules

Cell-surface remodelling during mammalian erythropoiesis.

Current evidence suggests that the major cell-surface modification occurring during mammalian erythropoiesis could be generated by two separate mechanisms: either selective loss of membrane proteins during enucleation or endocytosis at the subsequent reticulocyte and erythrocyte stages. The former idea was tested by collecting developing rabbit erythroid cells before and after the enucleation step and comparing their cell-surface protein composition via radiolabelling and electrophoresis. Few changes were observed. Our data thus lend support to the endocytosis mechanism.

Animals

Surface changes in chimpanzee sperm during epididymal transit.

Intact chimpanzee caput and cauda epididymal sperm, sperm cell lysates, and caput and cauda epididymal fluid were radiolabeled by enzymatic iodination with lactoperoxidase and Na125 I and were compared by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Caput epididymal sperm showed nine labeled macromolecular components of 90, 64, 56, 48, 38, 31, 20, 18 and 16 Kd and cauda epididymal sperm showed eleven macromolecular components of 90, 64, 55, 47, 42, 33, 27, 18, 17, 15 and 11 Kd. Six of the components labeled on caput sperm (90, 64, 56, 48, 18 and 16 Kd) were detected in equal amounts of cauda sperm and two (38 and 20 Kd) were detected at greatly reduced labeling intensities. In the cauda epididymidis, four new components (33, 27, 17 and 11 Kd) became prominent features of the sperm surface. Analysis of labeled caput and cauda sperm cell lysates resolved components distinct from those detected on sperm surfaces. Electrophoresis of caput epididymal fluid showed five labeled components of 66, 56, 47, 41 and 37 Kd, while electrophoresis of cauda epididymal fluid showed eight labeled components of 92, 66, 56, 48, 31, 27, 24 and 11 Kd. Three components (66, 56 and 47 Kd) were present in both caput and cauda fluid, two (41 and 37 Kd) in caput fluid only, and five (92, 31, 27, 24 and 11 Kd) in cauda fluid only. Components of 37 Kd were labeled in caput fluid and on caput sperm but not on cauda sperm, whereas components of 27 Kd and 11 Kd were labeled in cauda fluid and on cauda sperm but not on caput sperm. These data show that chimpanzee sperm undergo extensive surface modifications during epididymal maturation and that some of these modifications may be related to exogenous proteins/glycoproteins in epididymal fluids.

Animals

Mineral phases of calcium phosphate.

Many studies of calcium phosphate precipitation have been made using relaxation techniques in which the concentrations of the lattice ions are allowed to decrease as equilibrium is approached. Since the nature of the phases that form depend markedly on the solution composition, this decrease can lead to concomitant phase transformations during the crystallization experiments. The results of the present constant composition (CC) studies show that defect apatites may be formed under conditions of sustained supersaturation with a non-stoichiometric coefficient dependent on the pH of the growth medium. An important factor in analyzing these experiments is the initial surface modification and ion-exchange processes involving H+ and Ca2+ ions after inoculation of the supersaturated solutions. Thereafter, active growth sites may be eliminated as the crystals undergo lattice perfection. Transformation of dicalcium phosphate dihydrate to octacalcium phosphate, involving dissolution and subsequent nucleation and growth of the new phase, is also influenced by surface roughening of the initial phase. Typical inhibitors that reduce the rate of growth of seed crystals in supersaturated solutions may actually induce the nucleation of calcium phosphate phases when immobilized on inert surfaces. This may be a factor in the modulation of crystal growth in many biological systems.

Calcium Phosphates

The Binkhorst Medal Lecture. Biologic relationship between poly(methyl methacrylate) intraocular lenses and uveal tissue.

Implantation of a poly(methyl methacrylate) (PMMA) intraocular lens (IOL) after cataract surgery is associated with breakdown of the blood-ocular barrier and a foreign body inflammatory response. Although the material is biocompatible, it is not inert. The history of PMMA IOL implantation has reflected mechanical, chemical, and immunological interactions between the IOL and ocular tissues. The common link in these events is instability of the blood-ocular barrier. In this lecture, I discuss the nature of these interactions and present evidence that currently investigated methods of surface modification and capsular bag IOL implantation are effective in stabilizing the blood-ocular barrier.

Animals

Artificial heart valves: improved blood compatibility by PECVD a-SiC:H coating.

Implants are steadily increasing in importance as substitutions for body functions. With the present state of the art, the limitations of the application of cardiovascular implants are due to insufficient performance of biomaterials. Present research in this field is being concentrated on efforts to improve the thrombus resistance of conventional materials by coating with semiconducting materials to actively influence the electrochemical interaction between the condensed matter and blood proteins. Based on an electrochemical model of the interaction of fibrinogen with an artificial surface and the resulting requirements for improving hemocompatibility, a coating of amorphous hydrogenated silicon carbide deposited by plasma-enhanced chemical vapor deposition (PECVD) is presently under evaluation as a special coating material for cardiovascular prostheses and is herein described. In particular, first results are published concerning the optimum deposition parameters in the PECVD process and cell culture tests. Experimental results of comparative partial thromboplastin time studies serve the purpose of proving the validity of the electrochemical reaction model referring the hemocompatibility of implantable materials to their semiconducting surface properties. The aim of this article is to demonstrate a feasible method for an antithrombogenic surface modification based on doped amorphous silicon carbide films that is in full conformance to the above mentioned model.

Biocompatible Materials

Platelet adhesion and contact activation time tests on HEMA coated cellulose acetate membranes.

Surface modification of cellulose acetate dialysis membranes was carried out by 60Co radiation induced graft copolymerization of the hydrogel, hydroxyethyl methacrylate (HEMA). The degree of grafting was controlled by varying the HEMA monomer concentration in the grafting solution and the radiation dose. A continuous flow platelet adhesion test was designed which allows testing under conditions more closely approximating hemodialysis than other small scale in vitro tests. Platelet adhesion on treated membranes fell substantially with increasing surface HEMA concentration. The presence of HEMA on the membrane surface did not affect the membrane activated clotting times significantly.

Acrylates

Heparin surface modified intraocular lenses implanted in the monkey eye.

The biocompatibility of heparin surface modified poly(methyl methacrylate) intraocular lenses (IOLs) was evaluated in two experiments following implantation in the anterior and posterior eye chambers of adult cynomolgus monkeys. Throughout the study, large inflammatory cells and prominent pigment deposits were seen on the unmodified lenses, whereas the heparin surface modified IOLs remained almost free of precipitates. Similarly, fewer posterior synechias were observed in eyes implanted with surface modified IOLs in the posterior chamber than in eyes implanted with control lenses. Histopathological examination of enucleated eyes confirmed the clinical findings. These experiments strongly support the idea that surface modification with heparin is a useful way to reduce clinical complications following cataract surgery with IOL implantation.

Animals

In vitro biologic responses to native and surface-modified asbestos.

A comparative study was made of in vitro biologic responses to native chrysotile, amosite, and crocidolite and corresponding asbestos fibers whose surfaces were modified by metal oxides. Interferon induction by influenza virus was depressed by approximately 50% by all native asbestos whereas corresponding surface modified asbestos minimally affected this nonspecific cellular defense mechanism. The release of the cytoplasmic enzyme, lactate dehydrogenase (LDH), and lysosomal enzymes, beta-N-acetylglucosaminidase (beta-NAG) and beta-glucuronidase (beta-Gluc), by rat alveolar macrophages after exposure to either native or surface-modified asbestos (which is indicative of membrane damage) was monitored. Although both native and surface-modified asbestos induced significant leakage of LDH, generally, lesser amounts of the enzyme were released as a result of exposure to the latter than to native asbestos. Whereas all forms of native asbestos caused significant release of beta-NAG and beta-Gluc, leakage of these enzymes from macrophages exposed to surface-modified asbestos was minimal. In contrast to native asbestos which induced irritation of cell membranes, as indicated by hemolysis of sheep erythrocytes, surface-modified asbestos exhibited minimal hemolytic activity. The findings indicate that surface modification of different asbestos by metal oxides generally lessened the adverse effect of the native mineral on the aforementioned biologic entities.

Animals

Advanced biomaterials development from "natural products".

Natural substances and structures can serve increasingly well as biomedical products, given recent advances in understanding of requirements for biocompatibility and of methods for their preservation and surface tailoring. A successful example is the derivation of limb salvaging vessels, used in arterial reconstructive surgery, from human umbilical cords. There are numerous opportunities for additional product development from the umbilical cords' main ingredient, Wharton's gel, ranging from biolubricants to wound-healing aids. Major problems yet to be overcome with natural starting materials are their propensity for calcification and eventual biodeterioration. Surface modification of biomaterials to exhibit desired degrees of interaction with contacting viable tissues promises the greatest beneficial results. General principles of bioadhesion have broad applicability, predicting material behavior in environments as diverse as blood, saliva, and seawater.

Adhesiveness

Immunoselection of a human melanoma resistant to specific lysis by autologous tumor-infiltrating lymphocytes. Possible mechanisms for immunotherapeutic failures.

Intratumoral heterogeneity has been proposed as a possible basis for immunotherapeutic failure when tumor-specific agents such as tumor infiltrating lymphocytes (TIL) are employed for cancer therapy. To examine this issue, highly specific oligoclonal MHC class I-restricted cytolytic TIL grown in bulk culture from patient 397 were used to immunoselect a TIL-resistant variant tumor from the autologous cultured melanoma line 397-mel. Four cycles of immunoselection produced tumor 397-R4, a variant completely resistant to 397 TIL but not to allogeneic LAK cell lysis in 4-h 51Cr release assays. By flow microfluorometry analysis, this tumor variant had not lost MHC molecules, adhesion molecules, or a variety of tumor-associated Ag expressed by the parent tumor but showed decreased expression of many Ag examined. Failure of 397-R4 to cold target inhibit TIL lysis of 397-mel suggested that cell-surface modification was at least one mechanism causing TIL resistance. The inherent lysability of 397-R4 was equal to 397-mel, as confirmed by lectin-dependent cellular cytotoxicity, lysis by non-MHC restricted allogeneic TIL, and lysis by a second line of 397 TIL grown independently from tumor 397. Treatment of 397-R4 with IFN-alpha or IFN-gamma, +/- TNF-alpha for 72 h before cytolytic assays enhanced TIL lysis of this target slightly, and enhanced surface expression of MHC class I and II molecules and the adhesion molecule ICAM-1. The resistant phenotype of 397-R4 was evident in all clones of 397-R4 examined and has been maintained in serial culture for over 13 mo and through passage in nude mice, suggesting that such stable tumor variants may provide an in vivo escape mechanism from specific immune reagents such as TIL. Evolving patterns of TIL culture clonality over time, as well as the spontaneous emergence of different clones in two long term TIL cultures grown under identical conditions from the same source of cryopreserved tumor, were documented by analyzing TCR gene rearrangements and suggest that TIL from different culture passages or lines may be used to overcome resistant tumor subpopulations.

Animals

Ultrastructural histochemical alteration of the plasma membrane in chronic myelocytic leukemia.

Ultrastructural histochemical evaluation of the surface of normal human blood and bone marrow cells exposed to the pyroantimonate-osmium (PAO) reaction indicated the selective binding of pyroantimonate to certain cations (calcium, magnesium, and possibly sodium) associated with the plasma membrane of neutrophilic leukocytes and their developmental forms. Other leukocytes and their precursors did not exhibit plasma membrane PAO reactivity. The extent of surface binding was related to cell maturity, with maximal labeling evident in the mid and late promyelocytes; decreased binding occurred with subsequent maturation while myeloblasts were nonreactive. This study was initiated to ascertain if histochemical surface modifications of neutrophilic cells occur in certain myeloproliferative disorders. In this regard, we have been able to demonstrate a distinctive defect in the plasma membrane PAO binding characteristics of the leukemic cells in chronic myelocytic leukemia (CML). Limited binding of pyroantimonate to the plasma membrane of the leukemic cell series in four patients with CML contrasted with that of the normal granulocytic cell series and the neutrophilic cells seen in myelomonocytic leukemia (two patients), myelofibrosis (one patient), and acute myelocytic leukemia (three patients). Comparison of surface PAO reactivity of neutrophilic cells in all stages of maturation in two patients with CML in blast crisis revealed that, in the patient with 30% circulating blast cells, PAO reactivity was identical to that noted in CML, while in the patient with 80% circulating blast forms, the PAO reactivity of the maturing neutrophilic cells more nearly resembled that observed in neutrophilic cells from normal individuals. Many neutrophilic cells from patients with myelofibrosis and myelomonocytic leukemia and from one patient in severe blast crisis had large surface deposits of pyroantimonate considered to reflect increased membrane-associated reactive cation.

Antimony

Neoendothelial healing of modified EPTFE grafts.

Expanded polytetrafluoroethylene (EPTFE) grafts have poor neoendothelial healing characteristics and low patency rates after long-term implantation. The authors have shown that this is due to the low porosity of currently used EPTFE grafts (20-30 microns fibril length). The inner surface coated grafts made of long antithrombogenic material fibrils (40-60 microns) are desirable, especially for small diameter grafts. The authors have implanted these surface modified grafts (coated grafts) and noncoated grafts into abdominal arteries of rats and observed good patency rates, and the effects of surface modification of the grafts on stable endothelial tissue growth. The authors used four different kinds of grafts (fibril length, 20, 40, 60, and 90 microns, respectively) to investigate the effect of porosity. High porosity (long fibril length) grafts induce good neoendothelial healing and collagen production.

Animals

Effects of chemical modifications on the surface- and protein-binding properties of the light chain of human high molecular weight kininogen.

The light chain of kallikrein-cleaved human high molecular weight kininogen is solely responsible for its cofactor activity in blood clotting. Sequencing of the NH2-terminal region of the light chain reported herein identified the third kallikrein cleavage site of high molecular weight kininogen as Arg-437. The co-factor activity of high molecular weight kininogen consists of the capacity to bind to negatively charged surfaces and to factor XI or prekallikrein. Chemical modification of the histidines by either photooxidation or ethoxyformic anhydride affected the equivalent of 14-16 of 23 histidines available and resulted in over 90% loss in procoagulant activity. The modified protein had drastically reduced surface- and zinc-binding capacity, but it bound successfully to either factor XI or prekallikrein. In contrast, modification of two carboxyl groups, which led to approximately 80-90% loss of procoagulant activity, seriously compromised protein binding but left surface binding unaffected. All 3 tryptophans were modified at pH 4.0 with N-bromosuccinimide with a 70% reduction in procoagulant activity, but only 1 tryptophan was available for reaction at pH 7.35, resulting in a 50% loss in activity. Tryptophan modification at acidic pH affected protein binding but did not modify surface or zinc binding. Modification of both available tyrosine and 9 of 18 available lysine residues did not have a significant effect on the procoagulant activity of the light chain. These studies indicate that histidines participate in surface binding and that free carboxyl groups and tryptophan side chains are involved in binding of high molecular weight kininogen to other clotting factors.

Amino Acid Sequence

A new method for promoting adhesion between precious metal alloys and dental adhesives.

A new, simple method of modifying the adherend metal surface by a liquid Ga-Sn alloy (Adlloy) was applied to dental precious and base-metal alloys for adhesion with 4-META adhesive resin. Adhesions of 4-META resin to three other surface states--as-polished, oxidized at high temperature, and electroplated tin--were also performed for comparison with the adhesion on Adlloy-modified surfaces. Bond strength measurements were made, and the durability against water at the adhering interface was evaluated. The Adlloy-modified gold alloys (Type IV and 14 K) and silver-based alloys (Ag-Pd and Ag-Cu) showed not only high bond strengths but also excellent water durability at the adhesion interface. Surface modification by Adlloy, however, did not affect adhesion to Ag-In-Zn and base-metal (SUS, Co-Cr, and Ni-Cr) alloys. Adhesion to the tin-electroplated specimens was comparable with that to the Adlloy-modified specimens.

Acrylic Resins