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Damage to peripheral nerve tissue may cause loss of function in both the nerve and the targeted muscles it innervates. This study compared the repair capability of engineered nerve conduit (ENC), engineered fibroblast conduit (EFC), and autograft in a 10-mm tibial nerve gap. ENCs were fabricated utilizing primary fibroblasts and the nerve cells of rats on embryonic day 15 (E15). EFCs were fabricated utilizing primary fibroblasts only. Following a 12-week recovery, nerve repair was assessed by measuring contractile properties in the medial gastrocnemius muscle, distal motor nerve conduction velocity in the lateral gastrocnemius, and histology of muscle and nerve. The autografts, ENCs and EFCs reestablished 96%, 87% and 84% of native distal motor nerve conduction velocity in the lateral gastrocnemius, 100%, 44% and 44% of native specific force of medical gastrocnemius, and 63%, 61% and 67% of native medial gastrocnemius mass, respectively. Histology of the repaired nerve revealed large axons in the autograft, larger but fewer axons in the ENC repair, and many smaller axons in the EFC repair. Muscle histology revealed similar muscle fiber cross-sectional areas among autograft, ENC and EFC repairs. In conclusion, both ENCs and EFCs promoted nerve regeneration in a 10-mm tibial nerve gap repair, suggesting that the E15 rat nerve cells may not be necessary for nerve regeneration, and EFC alone can suffice for peripheral nerve injury repair.

Peripheral nerve injuries resulting from trauma or disease often necessitate surgical intervention. Although the gold standard for such repairs uses nerve autografts, alternatives that do not require invasive harvesting of autologous nerve tissues are currently being designed and evaluated. We previously established the use of scaffoldless engineered neural conduits (ENCs) fabricated from primary cells as one such alternative in sciatic nerve repair in rats [Baltich et al. (2010) In Vitro Cell Dev Biol Anim 46(5):438–444]. The present study establishes protocols for fabricating neural conduits from adipose-derived stem cells (ASCs) differentiated to either a fibroblast or neural lineage and co-cultured into a three-dimensional (3-D) scaffoldless tissue-ENC. Addition of ascorbic acid-2-phosphate and fibroblast growth factor (FGF)-2 to the medium induced and differentiated ASCs to a fibroblast lineage in more than 90% of the cell population, as confirmed by collagen I expression. ASC-differentiated fibroblasts formed monolayers, delaminated, and formed 3-D conduits. Neurospheres were formed by culturing ASCs on non-adherent surfaces in serum-free neurobasal medium with the addition of epidermal growth factor (EGF) and FGF-2. The addition of 10 ng EGF and 10 ng FGF-2 produced larger and more numerous neurospheres than treatments of lower EGF and FGF-2 concentrations. Subsequent differentiation to glial-like cells was confirmed by the expression of S100. ASC-derived fibroblast monolayers and neurospheres were co-cultured to fabricate a 3-D scaffoldless tissue-ENC. Their nerve-like structure and incorporation of glial-like cells, which would associate with regenerating axons, may make these novel, stem cell-derived neural conduits an efficacious technology for repairing critical gaps following peripheral nerve injury.

Accurate modeling of the impact of aerosols on climate requires a detailed understanding of the vertical distribution of aerosols. The Cloud–Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO) provides continuous high-resolution vertical profiles of aerosol properties on a near-global scale. Here the CALIPSO Vertical Feature Mask is used to document the three-dimensional (3D) frequency-of-occurrence distribution of aerosols over a broad region of the Atlantic Ocean, Africa, Europe, and the Americas. The 3D distributions illustrate the seasonal cycle in the zonal and meridional variability of the vertical profiles of mineral dust, biomass-burning smoke, polluted dust (external mixture of dust and smoke), and polluted continental aerosol, and also of their emissions sources and transport pathways. Four aerosol domains stand out in the product: dust over North Africa and the Middle East and smoke over southern Africa and South America. The transport pathways of African dust and smoke over the Atlantic are evident. The intertropical convergence zone (ITCZ) plays a clear role in limiting the southward transport of North African dust and northward transport of South African smoke. Dust and smoke are mixed in the ITCZ and consequently the highest probability of polluted dust is found there, even though the probabilities of dust and smoke in this region are relatively low. The mixing of dust and pollution has significant implications for cloud microphysical processes over a broad region of the Atlantic.

It has previously been shown that misfolded mutant Akita proinsulin in the endoplasmic reticulum engages directly in protein complexes either with nonmutant proinsulin or with "hProCpepGFP" (human proinsulin bearing emerald-GFP within the C-peptide), impairing the trafficking of these "bystander" proinsulin molecules (Liu, M., Hodish, I., Rhodes, C. J., and Arvan, P. (2007) Proc. Natl. Acad. Sci. U.S.A. 104, 15841-15846). Herein, we generated transgenic mice, which, in addition to expressing endogenous proinsulin, exhibit beta-cell-specific expression of hProCpepGFP via the Ins1 promoter. In these mice, hProCpepGFP protein levels are physiologically regulated, and hProCpepGFP is packaged and processed to CpepGFP that is co-stored in beta-secretory granules. Visualization of CpepGFP fluorescence provides a quantifiable measure of pancreatic islet insulin content that can be followed in live animals in states of health and disease. We examined loss of pancreatic insulin in hProCpepGFP transgenic mice mated to Akita mice that develop neonatal diabetes because of the expression of misfolded proinsulin. Loss of bystander insulin in Akita animals is detected initially as a block in CpepGFP/insulin production with intracellular accumulation of the precursor, followed ultimately by loss of pancreatic beta-cells. The data support that misfolded proinsulin perturbs bystander proinsulin in the endoplasmic reticulum, leading to beta-cell failure.

Nerve grafts are often required to replace tissue damaged by disease, surgery, or extensive trauma. Limitations such as graft availability, donor site morbidity, and immune rejection have led investigators to develop strategies to engineer nerve tissue. The goal of this study was to fabricate a scaffoldless three-dimensional (3D) nerve construct using a co-culture of fetal nerve cells with a fibroblast monolayer and allow the co-culture to remodel into a 3D construct with an external fibroblast layer and an internal core of interconnected neuronal cells. Primary fibroblasts were seeded on laminin-coated plates and allowed to form a confluent monolayer. Neural cells isolated from E-15 spinal cords were seeded on top of the fibroblast monolayer and allowed to form a networked monolayer across the monolayer of fibroblasts. Media shifts initiated contraction of the fibroblast monolayer and a remodeling of the co-culture into a 3D construct held statically in place by the two constraint pins. Immunohistochemistry using S100 (Schwann cell), beta3-tubulin, DAPI, and collagen I indicated an inner core of nerve cells surrounded by an external layer of fibroblasts. Conduction velocities of the 3D nerve and control (fibroblast-only) constructs were measured in vitro and compared to in vivo measures of neonatal sciatic nerve. The conduction velocities of the nerve constructs were comparable to 24-d-old neonatal nerve. The presence of Schwann cells and the ability to conduct neuronal signals in vitro suggest the scaffoldless 3D nerve constructs will be a viable option for nerve repair.