Membrane-seeded autologous chondrocytes: cell viability and characterization at surgery

The implantation of chondrocytes, seeded on matrices such as hyaluronic acid or collagen membranes, is a method that is being widely used for the treatment of chondral defects. The aim of the present study was to evaluate the distribution, viability and phenotype expression of the cells seeded on a collagen membrane just at the time of the implantation. Twelve patients who were suffering from articular cartilage lesions were treated by the MACI^® procedure. The residual part of each membrane was tested by colorimetric assay (MTT) and histochemical and ultrastructural analyses were carried out. In all of the samples a large number of viable cells, quite homogenously distributed, was detected. The cells expressed the markers of the differentiated hyaline chondrocytes. These data reassure in that the MACI procedure provides a suitable engineered tissue for cartilage repair, in line with the clinical evidences emerging in the literature.     

Program for the Prevention of Smoking in Secondary School Students

Background and ObjectiveSchool smoking prevention programs have never yielded the expected results. The aim of this study was to analyze the efficacy of an intensive smoking prevention program created by the educational community in which it was to be applied.Population and MethodA 3-year smoking prevention program was carried out among the students of Fuentesaúco Secondary School in Zamora, Spain. The Babilafuente Secondary School in Salamanca, Spain was the control group. The program included both prevention and treatment activities. The former were carried out in the school, in out-of-school situations, and in the community. The questionnaire of the European Smoking Prevention Framework Approach was used.ResultsA total of 417 students aged 12 to 17 years participated in the study. Of these, 54.4% belonged to the intervention group and 45.6% to the control group. Smokers represented 36.7% of the population. After the intervention smokers represented 40.1% of the Fuentesaúco students compared with 46.1% of the Babilafuente students, though the difference was not statistically significant. With respect to the cognitive determinants of smoking behavior, after the intervention significant differences in favor of the intervention group were only observed in the subjects’ perception of the behavior of their siblings, peers, and teachers.ConclusionsThe use of smoking prevention programs in schools should be reconsidered, and their evaluation should be based on educational rather than clinical criteria. Proposed changes in the program include decreasing its intensity, starting with students of an earlier age and seeking greater involvement of parents.     

Autologous implantation of chondrocytes on a solid collagen scaffold: clinical and histological outcomes after two years of follow-up

Abstract From our overall experience in 56 patients, we here report the treatment with matrix-induced autologous chondrocyte implantation (MACI) of 35 patients suffering from knee cartilage defects measuring about 4 cm², and followed for a minimum of 6 months. A total of 36 knees were treated (1 patient on both knees) and clinically observed for 22 months (in some cases for over 39 months), in accordance with a standardised protocol. Subjective parameters (pain, well-being, functional state, symptoms during specific activity) and objective outcomes (IKDC score and Lysholm and Tegner scores) were recorded. One or 2 years after implantation, some biopsies of the regenerated cartilage were histologically evaluated. The subjective parameters (VAS pain score, 2.80±1.49, p<0.0001; change vs. basal score, 2.72) promptly normalized after 1 month, as did the objective ones (IKDC score after 6 months, 1.53±0.59, p<0.0001; change vs. basal score, 1.78). Similar results were observed after the treatment of a femoropatellar kissing lesion. The three cartilage biopsies that were analysed from different patients showed a tissue positivity to immunohistochemical markers of hyaline cartilage. The conclusions of this preliminary analysis are that the clinical outcome and histological evaluation suggest that MACI is able to relieve pain and restore the functionality of the knee, and that the treatment appears capable of regenerating hyaline cartilage.     

Alpha-1-antichymotrypsin in renal biopsies

Alpha 1-Antichymotrypsin (alpha 1-AK) and alpha-1-antitrypsin (alpha 1-AT) represent a defense mechanism to protect the tissues from proteolytic enzyme activity. We studied the implication of alpha 1-AK and alpha 1-AT in glomeruli of patients with different nephropathies based on the analysis of 52 paraffin-embedded renal biopsies with alpha 1-AK and alpha 1-AT antisera. The results demonstrate an intense alpha 1-AK glomerular staining in renal biopsies from patients with minimal-change disease, while a minor staining of this protein was found in the other nephropathies. No significant evidence of alpha 1-AT deposits was observed in our cases. Our findings suggest that when alpha 1-AK is lacking in glomeruli the defense mechanisms against proteolytic enzymes may not be efficient enough to protect the glomerular structures and limit the damage. Since alpha 1-AK is a reactant of the acute phase of inflammation, it may be considered as a marker of activity for monocyte-macrophages in glomerular damage.     

Increased external tibial torsion in Osgood-Schlatter disease

We studied the relationship between Osgood-Schlatter disease and torsional abnormalities of the lower limb in 21 boys with this condition and 20 age- and sex-matched controls. 3 groups of knees (20 control knees, 21 symptomatic and 21 asymptomatic or less symptomatic knees) were subjected to clinical, radiographic and CT evaluation. We found no statistically significant differences between patients and controls, as regards femoral anteversion, patellar congruence angle, patellar tilt angle and anterior tibial tuberosity-trochlear groove distance, but the condylomalleolar angle and tibial torsion angle were greater in patients. We found no differences between symptomatic and asymptomatic or less symptomatic knees in any of the parameters. All the symptomatic knees were on the side preferentially involved in jumping and sprinting. This increase in external tibial torsion may play a role as a predisposing mechanical factor in the onset of Osgood-Schlatter disease in male athletes.     

Mitotic polyploidization in trophoblast giant cells of the alpaca

Genome multiplication is a typical feature of trophoblast giant cell (TGC) development in many species. Elevated nuclear DNA contents can be achieved by modified cell cycles with a complete lack of mitosis (endoreduplication) or with incomplete mitoses. The aim of this study is to characterize genome multiplication in the alpaca TGC. Placental tissues of gestation days 150, 264 and 347 (near term) and term placentae were processed for light microscopy and for transmission electron microscopy. Each TGC showed many nuclear profiles. Observation of serial sections revealed that TGCs are truly multinucleate with several highly lobulated nuclei. Feulgen staining showed that TGC nuclei have a higher DNA content than nuclei of other trophoblast cells. The number of argyrophilic nucleolar organizer regions (AgNORs) in nuclear profiles of TGC was between 15 and 100, while other trophoblast cells showed 1 or 2 AgNORs. Large multipolar mitotic figures with maximal diameters of 80 mum were observed in the alpaca placentas on gestation days 264 and 347. No cytokinesis was seen in TGC. The results show that the mode of genome multiplication in the alpaca TGC is mitotic polyploidization. Subsequent acytokinetic mitoses may lead to an accumulation of chromosomes and centrioles in TGC. With increasing ploidy levels, the shape of these polyploidizing mitoses becomes more irregular. The restitution of nuclei after these complex multipolar mitoses is likely to result in the irregular nuclear shape in TGC.