Morphological and ultrastructural characterization of neurospheres spontaneously generated in the culture from sheep ovarian cortical cells.

Neurospheres (NS) derived from adult stem cells of non-neural tissues represent a promising source of neural stem cells (NSCs) and neural progenitor cells (NPCs) for autologous cell therapy. Knowing the fine structure of NS cells is essential for characterizing them during differentiation or oncogenic transformation. NS are generated by culturing ovarian cortical cells (OCCs) under specific conditions. To establish whether these OCCs exhibited a similar morphophenotype as those from the central nervous system (CNS) reported in the literature, sheep OCCs were cultured for 21 days to generate NS. Expression levels of pluripotency (Nanog, octamer-binding transcription factor 4 [Oct4], and SRY-box transcription factor 2 [Sox2]) and NSCs/NPCs (nestin, paired box 6 [Pax6], and p75 neurotrophin receptor [P75NTR]) transcripts were analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), the NSC/NPC antigens were immunolocalized, and structural and ultrastructural analyses were performed in OCC-NS on Days 10, 15, and 21 in culture. Spheroids expressed transcripts and antigens of pluripotency as well as NSCs/NPCs. Cells were arranged into an inner core, with frequent apoptotic and degenerative events, and a peripheral epithelial-like cover with abundant cytoplasmic organelles, apical microvilli, and filament bundles of cytoskeleton elements. Adherens junctions and apical tight and lateral loose interdigitations were found in peripheral cells that eventually lost apical-basal polarization, which might indicate their disengaging/aggregating from/to the NS. We can conclude that OCC-NS shares the most structural and ultrastructural characteristics with CNS-NS.

Short-term outcomes and complications of 65 cases of porous TTA with flange: a prospective clinical study in dogs.

BACKGROUND: Cranial cruciate ligament rupture (CrCLR) is the most common orthopaedic cause of lameness in the hind limb in dogs. Many surgical treatments have been described, but tibial tuberosity advancement (TTA) is one of the most commonly used today. Since it was first described, TTA has evolved to reduce major complications and to arrest the progression of osteoarthrosis. The aim of this study was to assess a surgical technique called Porous TTA with flange prospectively. This study was performed in 61 dogs that underwent 65 Porous TTA with flange procedures, to validate it as an alternative CrCLR treatment. Complications and clinical outcomes (pain, lameness, weight bearing, flexion, extension, crepitation and atrophy) were reported over 3 months, i.e. at 3, 6 and 12 weeks postoperatively. RESULTS: The results showed a positive clinical outcome, a minor complication rate of 47.69% at the first review 3 weeks postoperatively, 10.77% at the second one (6 weeks after the surgery) and 4% at the third one (at 12 weeks). Major complications were observed only at the last review, with one case that had an infection requiring implant removal; this represented 1.5% of cases. Variables evaluated for a relationship with complication scores and improvement were body condition score, sex, age, breed, body weight, breed size, side of the affected limb, traumatic anamnesis and time of lameness before surgery. No relationship was detected. CONCLUSIONS: Clinical outcomes and complications show that Porous TTA with flange is an efficient alternative for surgical treatment of CrCLR in dogs.