An arthroscopic approach for the treatment of osteochondral focal defects with cell-free and cell-loaded PLGA scaffolds in sheep.

Osteochondral injuries are common in humans and are relatively difficult to manage with current treatment options. The combination of novel biomaterials and expanded progenitor or stem cells provides a source of therapeutic and immunologically compatible medicines that can be used in regenerative medicine. However, such new medicinal products need to be tested in translational animal models using the intended route of administration in humans and the intended delivery device. In this study, we evaluated the feasibility of an arthroscopic approach for the implantation of biocompatible copolymeric poly-D,L-lactide-co-glycolide (PLGA) scaffolds in an ovine preclinical model of knee osteochondral defects. Moreover this procedure was further tested using ex vivo expanded autologous chondrocytes derived from cartilaginous tissue, which were loaded in PLGA scaffolds and their potential to generate hyaline cartilage was evaluated. All scaffolds were successfully implanted arthroscopically and the clinical evolution of the animals was followed by non invasive MRI techniques, similar to the standard in human clinical practice. No clinical complications occurred after the transplantation procedures in any of the animals. Interestingly, the macroscopic evaluation demonstrated significant improvement after treatment with scaffolds loaded with cells compared to untreated controls.

Separation between the digestive and the respiratory lumina during the human embryonic period: morphometric study along the tracheo-oesophageal septum.

An isolated tracheo-oesophageal fistula could be caused by close proximity of the epithelia of both organs (O'Rahilly & Müller, 1984; Kluth et al. 1987) at certain embryonic stages, the most frequent location being the tracheal bifurcation. Thus the relative position and degree of separation between the digestive and the respiratory tubes throughout their development may be relevant to the origin of this anomaly. The aim of this study was to analyse along the different segments of the tracheo-oesophageal septum (TES) where the closest relationship between both lumina occurred and what degree of separation was present at each segment. Computer imaging techniques were applied on cross sections of a graded series of normal human embryos (Carnegie stages (CS) 13-23). In addition, the differentiation of the primitive TES was also studied (from CS 12) by light microscopy. Between CS 13 and 16 both tubes tended to separate (phase of separation), principally at the proximal segments of the laryngopharyngeal and the tracheo-oesophageal portions of the TES. During this phase the separation between the trachea and oesophagus was wider than between the larynx and pharynx. From CS 17 to CS 23 the digestive and respiratory lumina reached their widest separation at different levels of the laryngopharyngeal portion. Below these levels they tended to come closer together, principally at the proximal segment of the tracheo-oesophageal portion, but also at the distal part of the laryngopharyngeal portion. During this phase of approximation they reached their closest relationship at the proximal (CS 17) and the distal (from CS 18) segments of the tracheo-oesophageal portion. When finally the distal segment of the trachea (which includes the bifurcation) comes closest to the oesophagus, the coats of both organs have already undergone an appreciable differentiation. According to these observations, the origin of the most frequent isolated tracheo-oesophageal fistula at the bifurcation region could not be explained from the normal development of the TES.

Electrophysiological evaluation of spinal reflexes during epidural anesthesia in an experimental model.

We assessed the effects of epidural anesthesia with bupivacaine in the rat by serial recordings of spinal reflexes. The H wave from plantar muscles after electrical stimulation of the sciatic nerve evaluates a large nerve fiber spinal reflex arch. The extensor reflex response recorded from quadriceps muscle after stimulation of the contralateral tibial nerve assesses a reflex arch with small fiber afferents. After epidural injection of 0.2 mL of bupivacaine (0.25%, 0.5%, and 1.0% solutions) at the L5-L6 vertebral space, nociceptive, H, and extensor reflex responses were abolished within 1-3 min. Duration of complete blockade lasted 20-80 min, increasing with the anesthetic concentration, and complete recovery occurred after an additional period of 30-40 min. The responses recovered to amplitudes similar to preanesthesia controls, indicating that there was no damage to the nervous system. This study shows that electrophysiological recording and quantitation of nerve reflex responses is a useful and accurate method to evaluate the efficacy of local anesthetic agents.