Slow-release Vehicle Research Articles

Phytantriol phase behaviour in choline chloride urea and water mixtures.

Deep eutectic solvents (DES) are tailorable non-aqueous solvents with promising properties for a range of applications, from industrial dissolution of plant products to biomedicine. They are mixtures of hydrogen bond donors and acceptors with low melting points that can be tailored to specific applications, and many support the self-assembly of amphiphilic molecules into lyotropic liquid crystal phases. Self-assembled lipid structures have potential for numerous applications, including drug delivery. These ordered structures can act as carriers, slow-release vehicles, or microreactors. Lipid self-assembly in non-aqueous solvents, such as deep eutectic solvents, is important for applications at extreme temperatures, or involving water-insoluble or water sensitive components. However, lipid self-assembly in these solvents remains largely unexplored. In this paper, we have examined the self-assembly of phytantriol, a non-ionic lipid, at 10 and 30 wt% in the deep eutectic solvent choline chloride:urea, with and without water. Self-assembly was assessed using small angle X-ray scattering and cross polarised optical microscopy at temperatures from 25-66 °C. We found that pure choline chloride:urea supports a Pn3m cubic phase similar to that formed in water. However, mixtures of the DES with water resulted in phytantriol forming an inverse hexagonal phase and influenced the phase transition temperatures. These results demonstrate that choline chloride:urea can support diverse phase behaviour, and also provides a mechanism for tailoring the phase for particular applications simply by controlling the amount of water in the solvent. In the future this could lead to methods of triggered release of drugs and biomolecules by the simple addition of water which could be critical for drug delivery applications.

Device implant based on poly (lactic acid) with vitamin E for vaccine delivery system in Tilapia: Study for biocompatibility and biodegradation

The use of Poly (lactic acid) (PLA) as a slow-release vehicle for vaccines has attracted the attention of researchers, since its insertion improves the uptake of them, and reduces side effects or by stimulating recruited defense cells, assisting immunity without the need for booster vaccine doses. Seeking to develop new strategies for the administration of drugs and vaccines in aquaculture, we evaluated the biocompatibility and biodegradation of polymeric PLA devices and PLA plus vitamin E devices, implanted through subcutaneous (SC) and intraperitoneal (IP) routes in Nile tilapia. To carry out this study, 84 male tilapia (initial 243.82 ± 56.74 g; final 400.71 ± 100.54 g) were randomly distributed in 3 tanks (n = 28 fish per treatment/tank). The devices were prepared in two formulations: neat PLA (containing 100% PLA) and PLAVE (PLA plus vitamin E) implanted using a commercial AnimalTag® applicator, and non-implanted fish (control). Fish were sampled 15, 30, 60, and 120 days post-implantation (DPI). Blood analysis was used to access blood cells and blood smear for differential leucocytes count. Serum biochemistry to evaluated changes in serum proteins and glycemia. Histopathological investigation using hematoxylin-eosin (H&E) was used to assess polymer-tissue interaction. Histochemistry and immunohistochemistry was used to detection immune cells and phagocytes in capsule, and analyses of melanomacrophage centers (MMCs) to morphometric evaluation and percentage amount of melanin, hemosiderin and lipofucsin pigments. Histopathological study revealed an increase of capsular formation and inflammatory cell infiltration in PLAVE-implanted tilapia through SC route (15 DPI). Tilapia implanted with PLAVE and PLA (SC) presented mast cells and eosinophilic granular cells during 15, 30, and 60 DPI, with a decrease in these cells in the fibrous capsule around the polymer at 120 DPI. PLAVE implanted tilapia SC at 60 DPI showed significantly phagocytosis points than other groups. Phagocytic cells (F4/80+) were observed near to biopolymers in phagocytosis sites. Lipofuscin at 120 DPI in spleen melanomacrophage centers were significantly high in PLAVE implanted tilapias when compared to fish with PLA implants and control. The serum biochemical study of tilapia did not reveal changes in cytotoxicity and liver function in implanted fish. The absence of side effects in hematological and biochemical findings, including the absence of mortality after device implantation, proves its clinical safety. PLA implants in tilapia have demonstrated biocompatibility, biodegradation, clinical safety, and excellent evolution of foreign body inflammatory responses.

Alginate microencapsulation of an attenuated O-antigen mutant of Francisella tularensis LVS as a model for a vaccine delivery vehicle.

Francisella tularensis is the etiologic agent of tularemia and a Tier I Select Agent. Subspecies tularensis (Type A) is the most virulent of the four subspecies and inhalation of as few as 10 cells can cause severe disease in humans. Due to its niche as a facultative intracellular pathogen, a successful tularemia vaccine must induce a robust cellular immune response, which is best achieved by a live, attenuated strain. F. tularensis strains lacking lipopolysaccharide (LPS) O-antigen are highly attenuated, but do not persist in the host long enough to induce protective immunity. Increasing the persistence of an O-antigen mutant may help stimulate protective immunity. Alginate encapsulation is frequently used with probiotics to increase persistence of bacteria within the gastrointestinal system, and was used to encapsulate the highly attenuated LVS O-antigen mutant WbtIG191V. Encapsulation with alginate followed by a poly-L-lysine/alginate coating increased survival of WbtIG191V in complement-active serum. In addition, BALB/c mice immunized intraperitoneally with encapsulated WbtIG191V combined with purified LPS survived longer than mock-immunized mice following intranasal challenge. Alginate encapsulation of the bacteria also increased antibody titers compared to non-encapsulated bacteria. These data suggest that alginate encapsulation provides a slow-release vehicle for bacterial deposits, as evidenced by the increased antibody titer and increased persistence in serum compared to freely suspended cells. Survival of mice against high-dose intranasal challenge with the LVS wildtype was similar between mice immunized within alginate capsules or with LVS, possibly due to the low number of animals used, but bacterial loads in the liver and spleen were the lowest in mice immunized with WbtIG191V and LPS in beads. However, an analysis of the immune response of surviving mice indicated that those vaccinated with the alginate vehicle upregulated cell-mediated immune pathways to a lesser extent than LVS-vaccinated mice. In summary, this vehicle, as formulated, may be more effective for pathogens that require predominately antibody-mediated immunity.

Preparation of TTO/UF resin microcapsule via in situ polymerisation and modelling of its slow release

For slow release of tea tree oil (TTO), TTO were encapsulated by urea–formaldehyde (UF) resin via in situ polymerisation. The effects of curing time and drying condition on particle size, and TTO loading of the TTO/UF microcapsules were investigated. The results indicated that TTO/UF resin microcapsules with curing time of 80 min had narrow size distribution and good wall cover. Drying at ambient was better to maintain the TTO content than drying at oven. The loading of TTO with curing time of 80 min can be up to 45 wt.% of the mass-proportion to the prepared microcapsules, and more than 90 wt.% of the loaded TTO could be sustainably released in about 5 days. Moreover, the release kinetics of TTO/UF microcapsules was well described by Ritger-Peppas model, revealing non-Fickian diffusion. Promisingly, TTO/UF microcapsules with good stability can be used as a slow release vehicle for antibacterial application.

Eco-friendly smart hydrogels for soil conditioning and sustain release fertilizer

In the agricultural industry, biomass waste generated along with the production of food, is transformed into value-added products. Also, the demand for the slow or control release fertilizer is increasing to reduce the cost of maintaining the agricultural land. These issues are tackled by converting biomass waste to a slow fertilizer release vehicle and biodegradable water retention material that could reduce the cost for fertilizer application and simultaneously preserved the soil from dryness. A series of carboxymethyl sago pulp (CMSP) hydrogels were prepared by electron beam irradiation. Slow release systems were evaluated by loading potassium nitrate (KNO3) and ammonium nitrate (NH4NO3) into the CMSP hydrogels. These CMSP hydrogels endow slow release property, biodegradability and a soil moisture preservation property that could find greener application in agricultural industries. The results indicate that CMSP hydrogels have a great potential as an agent for slow release of fertilizers.

Enzyme- and gene-based approaches for developing methanogen-specific compounds to control ruminant methane emissions: a review

Methane emissions from ruminants are of worldwide concern due to their potential to adversely affect climate patterns. Methane emissions can be mitigated in several ways, including dietary manipulation, the use of alternative hydrogen sinks, and by the direct inhibition of methanogens. In the present review, we summarise and emphasise studies where defined chemically synthesised compounds have been used to mitigate ruminant methane emissions by direct targeting of methanogens and discuss the future potential of such inhibitors. We also discuss experiments, where methanogen-specific enzymes and pure cultures of methanobacterial species have been used to aid development of inhibitors. Application of certain compounds can result in dramatic reductions of methane emissions from ruminant livestock, demonstrating ‘proof of principle’ of chemical inhibitors of methanogenesis. More recently, genome sequencing of rumen methanogens has enabled an in-depth analysis of the enzymatic pathways required for methane formation. Chemogenomic methods, similar to those used in the fight against cancer and infectious diseases, can now be used to specifically target a pathway or enzyme in rumen methanogens. However, few rumen methanogen enzymes have been structurally or biochemically characterised. Any compound, whether natural or man-made, that is used as a mitigation strategy will need to be non-toxic to the host animal (and humans), cost-effective, environmentally friendly, and not accumulate in host tissues or milk products. Chemically synthesised inhibitors offer potentially significant advantages, including high levels of sustained inhibition, the ability to be easily and rapidly produced for global markets, and have the potential to be incorporated into slow-release vehicles for grazing animals.

VEGF Gene Transfer to the Utero-Placental Circulation of Pregnant Guinea Pigs to Enhance Fetal Growth.

In this chapter, we describe a novel method of adenoviral gene transfer to the uterine and radial arteries of pregnant guinea pigs to improve fetal growth. Adenoviruses encoding VEGF-A165 or a reporter gene β-galactosidase were reconstituted in pluronic gel and applied topically to the exposed uterine and radial arteries following laparotomy. Pluronic gel is a thermosensitive gel that is liquid at 4 °C, but becomes solid as soon as it comes in contact with body temperature. It thereby acts as a slow-release vehicle for viral vectors to the target tissue and also facilitates closer contact of the viruses with the host tissue. Our studies have shown that adenoviral gene delivery using pluronic gel resulted in the highest transduction efficiency compared to intra-arterial administration or external administration in PBS as a vehicle, when measured by X-gal staining, immunohistochemistry, or Western blotting.

Analysis of montmorillonite clay as a vehicle in platinum anticancer drug delivery

As a proof-of-concept study, the platinum anticancer complex [(1,10-phenanthroline)(1S,2S-diaminocyclohexane)platinum(II)]chloride, PHENSS, was loaded into montmorillonite (MMT) clay to evaluate its utility as a drug delivery vehicle. Loading is complete within one hour and the total amount of PHENSS that can be loaded into the clay is based on the PHENSS solution concentration in which the MMT is suspended. From a PHENSS solution concentration of 30mM, a maximum loading of 0.257mmol per gram of MMT can be achieved. The pH of the solution also has an effect with a solution pH of 6 giving maximum loading of PHENSS. Metal complex release from the MMT was examined using the dialysis bag and dispersion methods. PHENSS is incompletely released from MMT; after 4h just 47% has been released from the clay using the dialysis method and 30% using the dispersion method. The release is also very fast with a half-life of just 10–16min. The MMT was shown to have a negative effect on the in vitro cytotoxicity of PHENSS in the human breast cancer cell lines MCF-7 and MDA-MB-231, presumably due to the incomplete release of the metal complex from the clay. Overall the results demonstrate that MMT is not a suitable slow release vehicle for PHENSS, although it may still be of use to other platinum complexes and drugs.

IgG-mediated anaphylaxis to a synthetic long peptide vaccine containing a B cell epitope can be avoided by slow-release formulation.

Synthetic long peptides (SLP) are a promising vaccine modality to induce therapeutic T cell responses in patients with chronic infections and tumors. We studied different vaccine formulations in mice using SLP derived from carcinoembryonic Ag. We discovered that one of the SLP contains a linear Ab epitope in combination with a CD4 epitope. Repeated vaccination with this carcinoembryonic Ag SLP in mice shows improved T cell responses and simultaneously induced high titers of peptide-specific Abs. These Abs resulted in unexpected anaphylaxis after a third or subsequent vaccinations with the SLP when formulated in saline. Administration of low SLP doses in the slow-release vehicle IFA prevented the anaphylaxis after repeated vaccination. This study underscores both the immunogenicity of SLP vaccination, for inducing T cell as well as B cell responses, and the necessity of safe administration routes.

(483) - Efficacy and Safety of Sustained Release of Vancomycin Through Fibrin Glue Against Local Prosthesis Infection By Methicillin-resistant Staphylococcus aureus

Artificial device infection especially with methicillin-resistant Staphylococcus aureus (MRSA) often becomes critical in cardiovascular surgery. Topical antibiotic administration has been applied empirically; however, no comprehensive data exist showing long-lasting effects and safety of local antibiotic usage. We assessed fibrin glue (FG) as a slow-release vehicle for vancomycin (VCM) against local MRSA infection by means of animal experiments.

Long-term Treatment of Insulin-insensitive Mares with Cabergoline: Effects on Prolactin and Melanocyte Stimulating Hormone Responses to Sulpiride and on Indices of Insulin Sensitivity

The main experiment assessed whether the inhibitory effects of the dopamine agonist, cabergoline, on prolactin and α-melanocyte stimulating hormone (MSH) concentrations would persist throughout a longer-term administration (65 days). The possible effect of cabergoline on insulin sensitivity was also studied. Ten mares known to be insulin insensitive were allotted to two groups (treated vs. control). An insulin challenge, a glucose tolerance test, and a sulpiride challenge were administered before treatment. On day 0, treated mares (n = 5) received an injection of 5 mg cabergoline in slow-release vehicle; control mares (n = 5) received an equivalent vehicle injection. Injections were repeated every 10 days for a total of seven injections. Sulpiride challenges were done 1 day before each cabergoline treatment to assess possible refractoriness to the treatment. Behavior and hair coat density were also monitored. Plasma prolactin was suppressed (P < .01) to undetectable levels in mares receiving cabergoline; control mares had robust prolactin responses to each sulpiride injection. There was no indication of refractoriness to cabergoline over time. Plasma MSH concentrations after sulpiride were also suppressed (P < .05) by cabergoline. After treatment, neither the glucose response to insulin nor the insulin response to glucose differed (P > .1) between groups. No behavioral changes were noted because of treatment. Weight of hair samples indicated that cabergoline perturbed (P < .05) winter coat growth. It is concluded that 5 mg of cabergoline in slow-release vehicle administered every 10 days is an effective way of delivering dopaminergic activity to mares that results in no noticeable detrimental effects and no refractoriness to the drug.

Comprehensive evaluation of fibrin glue as a local drug-delivery system—efficacy and safety of sustained release of vancomycin by fibrin glue against local methicillin-resistant Staphylococcus aureus infection

Infection of prosthetic grafts, especially with methicillin-resistant Staphylococcus aureus (MRSA), often becomes critical. Although topical administration of antibiotics has been applied empirically, no comprehensive data are available showing long-lasting effects and safety of local antibiotic usage. By means of animal experiments we assessed fibrin glue (FG) as a slow-release vehicle for vancomycin (VCM) against local MRSA infection. Preliminary in-vitro experiments were performed to confirm that the FG-VCM mixture maintained viscosity as a sealant and led to slow-release of VCM. We next created a subcutaneous pocket in a rodent back and implanted a 1 cm (2) woven graft with 1 ml FG alone, or with serial concentrations of VCM (0-120 mg/ml. n = 3 for each group). MRSA of 1 × 10(7) colony-forming units (CFU) was injected into the pocket after wound closure. The graft was explanted 7 days later and was submitted for culture ("Culture-graft"). Blood samples were obtained for regular blood work, serum VCM concentration measurements, and blood culture ("Culture-blood"). The pocket tissue was also submitted for measurement of local VCM concentration. There was a remarkable infectious response in the group without vancomycin; however, no other groups developed any sign of infection. "Culture-graft" resulted in MRSA growth for V0 only. "Culture-blood" was negative in all groups, and only minimal serum concentrations of vancomycin were detected. One-dose topical administration of VCM via FG was effective against localized MRSA graft infection without systemic VCM administration. Topical administration of antibiotics may help treat difficult graft infections and reduce systemic use of potent antibiotics.

Inhibitory Effects of Pergolide and Cabergoline Formulations on Daily Plasma Prolactin Concentrations in Geldings and on the Daily Prolactin Responses to a Small Dose of Sulpiride in Mares

Two experiments were conducted to assess the efficacy and duration of action of two dopaminergic compounds, pergolide and cabergoline, on daily prolactin secretion in geldings and on prolactin responses to a small dose of sulpiride over 10 days. In the first experiment, oral administration of 2 mg of pergolide was compared to a single injection of 2 mg of pergolide in a slow-release vehicle and a single injection of 5 mg of cabergoline in slow-release vehicle. Controls received vehicle only. All drug treatments reduced (P < .05) prolactin concentrations relative to that in controls but differed substantially in duration of action (oral pergolide approximately 6 hours or less, injected pergolide 6 to 24 hours, and injected cabergoline at least 6 days). In the second experiment, repeated small doses of sulpiride (2 μg/kg of body weight intravenously) were used to stimulate prolactin release in mares, and the ability of seven daily injections of pergolide (2 mg each) and a single injection of cabergoline (5 mg) in slow-release vehicle to suppress this release were compared. Control mares receiving vehicle injections had robust prolactin responses to the sulpiride injections on all days of injection (days 1, 0, 1, 2, 3, 4, 6, 8, and 10 relative to treatment). Prolactin responses were muted (P < .05) by pergolide and cabergoline treatments on the first day of injection (day 0, 30 min after treatment) and were basically absent on days 1 to 8. The single injection of cabergoline continued to be suppressive through day 10, whereas mares previously treated with pergolide (through day 6) had begun to recover a prolactin response by day 10. We conclude that either daily 2-mg pergolide injections in slow-release vehicle or a single injection of 5 mg of cabergoline in slow-release vehicle is an effective way to apply dopaminergic activity to horses for approximately 7 to 10 days and may have application in the treatment of pituitary pars intermedia dysfunction in affected horses.

Relaxin Does Not Rescue Coronal Suture Fusion in Craniosynostotic Rabbits

Craniosynostosis affects 1 in 2000 to 3000 live births and may result in craniofacial and neural growth disturbances. Histological data have shown that thick collagenous bundles are present in the sutural ligament, which may tether the osteogenic fronts, resulting in premature fusion. The hormone relaxin has been shown to disrupt collagen fiber organization, possibly preventing craniosynostosis by relaxing the sutural ligament and allowing osteogenic fronts to separate normally and stay patent. This study tested this hypothesis with a rabbit model of delayed-onset coronal suture synostosis. A total of 18 New Zealand White rabbits with craniosynostosis were randomly assigned to one of three groups: sham control, protein control (BSA), relaxin treatment. After initial diagnosis, sham surgery, BSA, or relaxin was delivered to the fusing coronal suture in a slow-release (56-day) collagen vehicle. Longitudinal radiographs and body weights were collected at 10, 25, 42, and 84 days of age, and sutures were harvested for histology. Relaxin-treated animals had more disorganized intrasuture content than control groups. These specimens also appeared to have relatively wider sutures ectocranially. There were no significant differences in relaxin-treated animals for all craniofacial growth measures, or suture separation compared with controls. These data do not support our initial hypothesis that the use of relaxin may rescue sutures destined to undergo premature suture fusion. These findings suggest that collagen fiber arrangement may not be important for suture fusion. This protein therapy would not be clinically useful for craniosynostosis.

Use of Channel Catfish, Ictalurus punctatus, Pituitary as a Spawning Aid: Influence of Seasons and Preparations on Efficacy

Channel catfish, Ictalurus punctatus, pituitary powder (CP) was prepared and administered to determine the effect of pituitary preparation, head selection criteria, and season of collection on the biological potency of this gonadotropic preparation. Pituitaries were harvested from three catfish processing plants throughout the months of December, February, March, and April to investigate the effect of season on CP potency. Gravid female channel catfish were injected intraperitoneally (IP) with priming (2 mg/kg) and resolving doses (8 mg/kg) of CP treatments, luteinizing hormone-releasing hormone analog (LHRHa), or saline. Pharmaceutical grade Carbopol 971P resin (CAR) was tested as a 1% aqueous dispersion in physiological saline and administered with CP (10 mg/kg) as a IP single injection at a final volume of 2 mL per kg as a slow-release vehicle. Individual females were hand-stripped, and eggs were fertilized with freshly harvested blue catfish, Ictalurus furcatus, spermatozoa. Pituitaries processed with either acetone or ethanol/acetone resulted in significantly different ovulation rates of 35 and 73%, respectively (P≤ 0.01). Ovulation success was not significantly different among CP and LHRHa treatment groups. Ovulation rates were not significantly different among treatment groups of CP collected in April (74%) or December (67%), or collected from male (69%), or small individuals (55%). The shortest latency period among CP treatment was from CP collected in December and from males at 39.1 ± 0.7 and 38.5 ± 0.4 h, respectively. The CAR vehicle treatment performed poorly resulting in an average ovulation rate of 19.0%, with 3.3% viable embryos at 36 h post-fertilization. Pituitaries collected from channel catfish potentially represent a viable spawning aid for the production of hybrid channel × blue catfish.

Liposome−Hydrogel Bead Complexes Prepared via Biotin−Avidin Conjugation

Liposomes immobilized onto polymeric hydrogel microbeads have potential advantages both in tissue engineering applications and as drug delivery vehicles. Here we demonstrate, quantify, and optimize lipid vesicle binding to polymeric hydrogel microbeads via the avidin-biotin conjugation system and characterize the stability of the resulting microgel-bound liposomes. Microgels consisting of a copolymer of N-isopropylacrylamide (NIPAM) and acrylic acid (AA), cross-linked with bis-acrylamide, that is, p(NIPAM-co-AA), were biotinylated using aqueous carbodiimide chemistry. Extruded liposomes consisting of 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) plus a small fraction of a biotin-derivatized phosphatidylethanolamine (B-PE) were saturated with avidin and allowed to bind to biotinylated hydrogel beads. Using a combination of fluorescence spectroscopy, quenching, and microscopy and 31P NMR static and magic angle spinning (MAS) spectroscopies, we demonstrate conditions for near-quantitative liposome binding to p(NIPAM-co-AA) microbeads and show that liposome fusion does not occur under such conditions, that the liposomes remain intact and impermeable when so bound, and that they can function as slow release vehicles for entrapped aqueous species.

Use of Carbopol resin for carp pituitary administration improves the fertilization percentage of northern pike (Esox lucius Linnaeus) eggs in commercial hatcheries

The efficiency of northern pike (Esox lucius Linnaeus) fry production via hormonal treatment of wintered broodstock is, in general, relatively low due to low egg fertilization percentage. It has been experimentally demonstrated that administration of acetone-dried carp pituitary extract in a slow-release vehicle of aqueous dispersion of Carbopol 971 P resin (CP) resulted in a higher mean fertilization percentage, possibly because the gradual hormone action could optimally control ovarian follicle maturation and ovulation. This new method of hormonal induction of ovulation was tested in a large-scale hatchery production of northern pike fry. In the present study, data on fertilization percentage collected between 2002 and 2006 at two prominent hatcheries in Hungary were analyzed. Administration of acetone-dried carp pituitary in a 2.5% CP vehicle resulted in higher fertilization (76.4 ± 7.7%; mean ± SD) compared to the saline vehicle group (59.3 ± 10.8%). The vehicle used did not affect the ovulation ratio. According to these results from large-scale production, the new method could be suitable to have wide application for induced breeding of northern pike.

Controlled Release of rhBMP-2 Loaded Poly(DL-lactic-co-glycolic Acid)/ Calcium Phosphate Cement Composites In Vivo

The release kinetics of recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded poly(DL-lactic-co-glycolic acid)/calcium phosphate cement (PLGA/Ca-P cement) composites were studied in vivo. RhBMP-2 was radiolabeled with 131I and entrapped within PLGA microparticles or adsorbed onto the microparticle surface. PLGA microparticles were prepared of high molecular weight (HMW) PLGA (weight average molecular weight [Mw] 49,100 ± 1,700) or low molecular weight (LMW) PLGA (Mw 5,900 ± 300) and used for preparation of 30:70 wt% PLGA/Ca-P cement composite discs. Release of 131I-rhBMP-2 loaded composites was assessed by scintigraphic imaging according to a 22 two-level full factorial design in the rat ectopic model during four weeks. In vivo release kinetics varied among formulations. All formulations showed slow release without initial burst, and displayed a linear release from 3 to 28 days. Release of LMW entrapped rhBMP-2 composites (1.7 ± 0.3%/day) was significantly faster than release from other formulations (p &lt; 0.01). After 28 days, retention within the composites was 65 ± 5%, 75 ± 4%, 50 ± 4% and 70 ± 6% of the initial rhBMP-2 for HMW entrapped, HMW adsorbed, LMW entrapped and LMW adsorbed rhBMP-2 composites, respectively. Release from the composite was probably slowed down by an interaction of rhBMP-2 and Ca-P cement after rhBMP-2 release from PLGA microparticles. We conclude that PLGA/Ca-P cement composites can be considered as sustained slow release vehicles and that the release and retention of rhBMP-2 can be modified according to the desired profile to a limited extent.

Controlled release of rhBMP-2 loaded poly( dl-lactic- co-glycolic acid)/calcium phosphate cement composites in vivo

The release kinetics of recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded poly( dl-lactic- co-glycolic acid)/calcium phosphate cement (PLGA/Ca–P cement) composites were studied in vivo. RhBMP-2 was radiolabeled with 131I and entrapped within PLGA microparticles or adsorbed onto the microparticle surface. PLGA microparticles were prepared of high molecular weight (HMW) PLGA (weight average molecular weight [ M w] 49,100 ± 1700) or low molecular weight (LMW) PLGA ( M w 5,900 ± 300) and used for preparation of 30 : 70 wt.% PLGA/Ca–P cement composite discs. Release of 131I–rhBMP-2 loaded composites was assessed by scintigraphic imaging according to a 2 2 two-level full factorial design in the rat ectopic model during four weeks. In vivo release kinetics varied among formulations. All formulations showed slow release without initial burst, and displayed a linear release from 3 to 28 days. Release of LMW entrapped rhBMP-2 composites (1.7 ± 0.3%/day) was significantly faster than release from other formulations ( p < 0.01). After 28 days, retention within the composites was 65 ± 5%, 75 ± 4%, 50 ± 4% and 70 ± 6% of the initial rhBMP-2 for HMW entrapped, HMW adsorbed, LMW entrapped and LMW adsorbed rhBMP-2 composites, respectively. Release from the composite was probably slowed down by an interaction of rhBMP-2 and Ca–P cement after rhBMP-2 release from PLGA microparticles. We conclude that PLGA/Ca–P cement composites can be considered as sustained slow release vehicles and that the release and retention of rhBMP-2 can be modified according to the desired profile to a limited extent.

Effects of pulsatile secretion of growth hormone (GH) on fat deposition in human GH transgenic rats

Growth hormone (GH) is an endocrine regulator of glucose and lipid metabolism as well as body growth. GH levels are decreased and a unique pulsatile secretory pattern becomes obvious after puberty particularly in males. Coincidentally with this, males tend to deposit body fat. Experimental and clinical evidence has accumulated that obesity is associated with a decrease in GH levels. A strain of transgenic rats has been generated with severe obesity but normal nose-to-tail length, which has low circulating GH levels without pulsatility (human growth hormone (hGH) transgenic rats). The present review mainly focuses on recent and current work analysing the relationship between the occurrence of obesity and low GH levels and/or the absence of GH pulsatility in this transgenic animal model. This model has elevated blood glucose, non-esterified fatty acid, insulin and leptin levels associated with hyperphagia, suggesting that these rats also carry insulin- and leptin-resistant characteristics. hGH transgenic rats were subjected to a pair-feeding treatment to normalize food intake and chronic GH replacement to normalize GH levels. While the pair-feeding for 8 weeks successfully suppressed body-weight gain, the fat pad : body weight ratio remained very similar to freely-eating control hGH transgenic rats, which indicates the hyperphagia is not the sole contributor to the excess fat accumulation in this model. However, continuous elevation of peripheral hGH levels (approximately 2-fold) for 8 weeks by means of a slow-release vehicle resulted in a significant decrease in the fat mass : body weight ratios by 30 %. This GH treatment altered neither food intake nor body-weight gain. Thus, two characteristic phenotypes observed in the hGH transgenic rats, hyperphagia and obesity, seem to be closely related to GH levels and GH secretory pattern. This relationship might be working in the regulation of changes in seasonal body composition in wild animals.