Optimizing the Bioavailability of Subcutaneously Administered Biotherapeutics Through Mechanochemical Drivers.

The subcutaneous route offers myriad benefits for the administration of biotherapeutics in both acute and chronic diseases, including convenience, cost effectiveness and the potential for automation through closed-loop systems. Recent advances in parenteral administration devices and the use of additives which enhance drug dispersion have generated substantial additional interest in IV to SQ switching studies. Designing pre-clinical and clinical studies using SQ mediated delivery however requires deep understanding of complex inter-related physiologies and transport pathways governing the interstitial matrix, vascular system and lymphatic channels. This expert review will highlight key structural features which contribute to transport and biodistribution in the subcutaneous space and also assess the impact of drug formulations. Based on the rapidly growing interest in the SQ delivery route, a number of potential areas for future development are highlighted, which are likely to allow continued evolution and innovation in this important area.

Influence of glial growth factor and Schwann cells in a bioresorbable guidance channel on peripheral nerve regeneration.

Using an established rat peripheral nerve regeneration model, we investigated the role of glial growth factor (GGF) in nerve regeneration in combination with a novel bioresorbable poly(lactic-co-glycolic) acid (PLGA) guide in vivo. Schwann cells, established from a 1-cm segment of excised rat sciatic nerve, were isolated and seeded onto nerve guides with or without GGF (n = 24/group). Living nerve guides were re-established in these animals, and nerve regeneration was assessed over a period of 12 weeks. Histological studies revealed a reduction in the total axon count and the number of myelinated axons in the presence of exogenously added Schwann cells compared to saline controls. In contrast, the addition of GGF alone enhanced the total number of axons and significantly increased the number of blood vessels. Although combining GGF with Schwann cells negated the enhanced numbers of axons and blood vessels seen with GGF alone, this combination resulted in the highest myelination index and the fastest conduction velocities recorded. The PLGA guide material did not trigger any histologically detectable host response and was permissive for nerve regeneration in this animal model. The results from this study demonstrate the potential utility of this guide in vivo and establish a promotional role for GGF in nerve regeneration.

Migration of schwann cells in peripheral-nerve regeneration.

Schwann cells play a central role in peripheral-nerve regeneration, in which it has been shown that the addition of exogenous Schwann cells enhances the temporal and spatial sequence of events observed in regeneration. In this study, the authors investigated the fate of exogenous cells in this process by using fluorescently tagged autogenous Schwann cells in an established rat model of peripheral-nerve regeneration. Tracking labeled cells over a 4-week period revealed early migration of Schwann cells into the proximal nerve segment, followed by a concentration of migrating Schwann cells, leading the proximal growth cone throughout the regenerative process. The early proximal distribution of labeled cells suggests active migration in response to nerve damage, with spatial localization at the center of the proximal nerve segment and not the epineural surface. These observations demonstrate an interaction of exogenous Schwann cells with intact nerve tissue in vivo and affirm their role in the directional growth of regenerating axons.

Hyaluronic acid enhances peripheral nerve regeneration in vivo.

Hyaluronic acid has been shown to enhance peripheral nerve regeneration in vitro. It has been proposed that, during the fibrin matrix phase of regeneration, hyaluronic acid organizes the extracellular matrix into a hydrated open lattice, thereby facilitating migration of the regenerating axons. Hyaluronic acid solutions and saline control solutions were injected into a nerve guide spanning a transected gap in the sciatic nerve of Sprague-Dawley rats (five in each group). Nerve conduction velocities were measured at 4 weeks by electromyography (EMG) before sacrifice of the animals. These studies demonstrated increased conduction velocities in the hyaluronic acid group compared with control animals (P = 0.006). After the animals were sacrificed, regenerated axon cables were quantified histologically, and axon branching was delineated by retrograde tracer analysis. In addition, the hyaluronic acid group showed an increase in myelinated axon counts at 4 weeks (P= 0.03). An increase in retrograde flow was demonstrated in the hyaluronic acid groups compared with animals receiving saline solution.

Effect of Schwann cells in the enhancement of peripheral-nerve regeneration.

Schwann cells appear to stimulate the early phases of axon regeneration. The reported study investigated whether nerve guides with Schwann-cell monolayers can help regenerating nerves span gaps larger than 1 cm. Schwann-cell cultures were established by resecting 1-cm segments of sciatic nerves of adult female Sprague-Dawley rats, establishing cell monolayers in 24-mm nerve guides, and then reinserting these "living guides" into 20-mm nerve gaps of the rats from which they were developed. Control groups had plain guides (no Schwann cells) inserted between the same 20-mm gaps. In the experimental group, resected nerves regrew progressively. At 8 weeks, regrowth had spanned the entire gap in 60 percent of the animals. Axon counts increased at each successive time point. Regeneration did begin to occur in the control group but by 8 weeks, those cables had atrophied. The experimental groups displayed more central connections and higher nerve conduction velocity. Explant organ cultures of rat sciatic nerve can be used to develop nonneural conduits with Schwann-cell monolayers. These living artificial nerve guides permit the spanning of gaps of at least 20 mm.

Surgical management of life-threatening epistaxis in Osler-Weber-Rendu disease.

Hereditary hemorrhagic telangiectasia is a systemic disorder of blood vessels. It often leads to severe epistaxis that is refractory to conventional therapy. Three patients with severe epistaxis caused by hereditary hemorrhagic telangiectasia unresponsive to laser coagulation and to septal dermoplasty and requiring multiple transfusions underwent extirpation of nasal mucosa through a rhinotomy approach, with a median forehead flap and split-thickness skin graft coverage of the mucosal defects over the floor and lateral walls of the nose. All patients had reduced frequency and severity of bleeding without any need for transfusions during follow-up periods of 6 months, 3 years, and 5 years, respectively. Forehead flaps resulted in minimal morbidity. Partial nasal obstruction and a forehead scar were the main complaints. Surgical technique, complications, and alternative therapy are discussed.

Continuous alteration of the internal milieu of a nerve-guide chamber using an osmotic pump and internal exhaust system.

The use of non-neural conduits to span gaps in regenerating peripheral nerves has been noted in the literature for many years. An intriguing addition to this so-called entubulation repair method is the continuous instillation of neuronotrophic or growth factors into the lumen of the guide to bathe the regenerating nerve. A model is presented which uses an osmotic pump that supplies a constant, reproducible amount of solution into the lumen of a nerve guide without disrupting the regeneration process.

Inside-out vein graft repair compared with nerve grafting for nerve regeneration in rats.

The inside-out vein graft is a vein conduit pulled through itself to invert the normal orientation and place the adventitial layer within the lumen of the conduit. Our study compares regeneration of peripheral nerves in the rat through two conduits: inside-out graft of the jugular vein and autogenous nerve graft. In 10 rats, the right jugular vein was harvested, turned inside out, and used to bridge a 10 mm defect created in the right sciatic nerve. The 10 mm nerve segment from the right was then used as a standard nerve graft to bridge a 10 mm gap created in the left sciatic nerve. Rats were sacrificed at 8 and 12 weeks. Regeneration on the inside-out vein graft side showed superior functional results (faster conduction velocities) and improved histological results (greater axon counts) compared with the nerve grafted side. We feel the adventitial surface of the wall of the vein promotes nerve regeneration by providing an environment rich with collagen, laminin, and Schwann cells and promotes increased vascularization of the new nerve.

Immunocytochemistry of skeletal muscle basal lamina grafts in nerve regeneration.

The influence on nerve regeneration of the extracellular matrix glycoprotein laminin was studied after sciatic nerve transection in 90 outbred Sprague-Dawley rats. Nerve regeneration through basal lamina grafts was comparable with regeneration through traditional nerve grafts across gaps up to 2.0 cm in length. True axonal regeneration rather than axonal branching was demonstrated by retrograde horseradish peroxidase labeling of nerve cables. Pretreatment of basal lamina grafts with antilaminin antibodies reduced the total number of regenerated axons by 90 percent with a significant decrease of nerve conduction velocity and a significant impairment of walking track patterns. The basement membrane glycoprotein laminin serves a critical role in the regeneration of peripheral nerves through basal lamina grafts.

Inside-out vein graft promotes improved nerve regeneration in rats.

Vein grafts have been used both experimentally and clinically to bridge gaps in peripheral nerves. This study describes a modification of the vein graft technique in which vein graft conduits are pulled inside-out before anastomosis with proximal and distal nerve stumps. This technique creates an autogenous vein conduit with the collagen-rich adventitial surface exposed to the regenerating axons. The inside-out technique is a fast and simple modification of the standard vein graft technique and produces an accelerated rate of nerve regeneration and significantly earlier myelination compared with the results obtained from the use of polyethylene nerve guides and standard vein graft conduits.

Common complications of wound healing. Prevention and management.

The culmination of any operation is a healed wound. Failure of a wound to heal increases time spent in the hospital and the expense, and may start a cascade of progressive complications. Thus, it is important to optimize as many factors as possible before operation and to have the knowledge and resources to deal with wound problems should they occur. This approach involves understanding basic wound healing with its multiple factors, including nutrition, and knowing how to deal with potential adverse factors in wound healing, such as chemotherapy, corticosteroids, and radiation.

Surgical management of ulnar artery aneurysms.

In a ten-year review (1978 to 1988), ten ulnar artery aneurysms in nine male patients were studied. Blunt trauma led to 70%, penetrating trauma to 20%, and 10% had no history of trauma. Three cases were seen as asymptomatic palmar masses with brief antecedent histories of 4 weeks or less. Seven patients with aneurysms had persistent vascular hand symptoms for 6 weeks or longer. All seven symptomatic lesions proved to be sources of emboli. Diagnostic arteriography was done in all cases. Intraoperative digital plethysmography aided in operative decisions regarding the necessity for microvascular reconstruction. Five aneurysms were resected with end-to-end ulnar artery microvascular repairs, four resected without repair, and a single case treated with long-term anticoagulants. Follow-up, averaging 40 months, showed uniform improvement in vascular symptoms, with no loss of jeopardized tissues. Ulnar artery aneurysms, well studied preoperatively and intraoperatively, can be treated successfully with selective microvascular reconstruction.

Erythrocyte membrane vesiculation and changes in membrane composition during storage in citrate-phosphate-dextrose-adenine-1.

Serial studies were made of the membranes of the erythrocytes and the vesicles shed during storage of blood in polyvinyl chloride containers for 35 days in citrate-phosphate-dextrose-adenine anticoagulant. Special precautions were taken to eliminate artifacts created by contaminating leukocytes, platelets and red blood cell ghosts. A total of 15.6% of the cholesterol and 5.2% of the phospholipids of the membranes was lost with no gross change in the gel electrophoretic patterns. The quantity of vesicles found in the supernatant plasma increased during storage and their membranes were characterized by the absence of spectrin, ankyrin, and periodic acid Schiff bands 2 and 3. The ratio of lipids to protein in the vesicles increased as they accumulated perhaps reflecting a rearrangement of the erythrocyte membrane constituents during prolonged maintenance at 4 degrees C.

Direct estimation of the frequency of human cytotoxic T lymphocytes and their precursors following in vitro allosensitization.

Cell mediated lympholysis (CML) has been proposed as an in vitro model of the rejection process that results from transplantation of allogeneic tissue. To date, the absolute frequencies of cytotoxic T lymphocytes (CTL) and their precursors (CTL.P) have not been directly estimated in man because of technical difficulties. Through optimizing the conditions for radiometric detection of 51Cr release and the attendant improvement in CML sensitivity, direct CTL frequency estimates have been determined in peripheral blood (PBL), spleen (SPL), and lymph nodes (LNC) after in vitro allostimulation using unrelated human cells and limiting dilution assays. The mean frequency of CTL generated from PBL is 1 in 826 cells (0.121% +/- 0.101%) which, from preliminary experiments, is significantly greater than that generated from either LNC or SPL (p less than 0.05). With restimulation of primed cells on day 10, the frequency of CTL generated from PBL was increased 400%. The CTL.P frequency (0.0064% +/- 0.0050%) was approximately 5% of the corresponding CTL frequency. The CTL.P frequencies were found to be minimal estimates as both accessory "filler" cells and T cell growth factors increased the level of detection of CTL.P an average of threefold. The limiting cell dilution assay as detailed in this report should be a powerful tool for defining the cellular requirements and related factors necessary for optimal induction of a CTL response and should provide the means for determination of the immunogenetic requirements and the allospecificity of human cytotoxic lymphocytes.