Application of tissue-engineered cartilage with BMP-7 gene to repair knee joint cartilage injury in rabbits.

Injured articular cartilage has a poor capacity for spontaneous healing. So far, satisfactory solution to this subsistent problem has not been found, but transgenic therapy may be a promising way. This study aims to evaluate the effectiveness of a tissue-engineered cartilage that was transfected with morphogenetic protein 7 (BMP 7) in repairing the cartilaginous defects of rabbit knee joints. Chondrocytes were transfected with BMP-7 gene (5 x 10(6) cells/ml), inoculated into the collagen-fibrin gel scaffolds, and cultured for 14 days. Then, the scaffolds were implanted onto the created defects (5.0 mm in diameter) in rabbits' knee joints. After 12 weeks, the rabbits were sacrificed and histological sections were evaluated using modified O'Driscoll cartilage scores; In situ hybridization and immunohistochemistry were performed to detect the expression of BMP-7 mRNA and BMP-7 at the implanted site while the content of DNA and GAG was determined as well. A better quality of repairs was observed at the 12th week after implantation when compared to the control group using histological analyses. The content of DNA and specific secretion of GAG in the treatment group is statistically significant different compared with the control group. Gene therapy may be a promising treatment method, but the novel therapy approach needs further studies with respect to a longer follow-up period.

The classification of posterior petrous meningiomas and its clinical significance.

This retrospective analysis of the clinical records of 42 patients was used to study the clinical significance of a classification system for posterior petrous meningiomas. According to clinical manifestations and radiological images, posterior petrous meningiomas were classified into three types: type I (cerebella type; 12 patients), tumours involved and compressed the cerebellum; type II (cranial nerve type; 16 patients), tumours involved the cranial nerves; and type III (combined type; 14 patients), tumours involved more than one structure such as the cerebellum, cranial nerves and the brain stem. All patients underwent microneurosurgery and the total resection rate was 90%. It was more difficult totally to resect type II and III tumours than type I tumours and the post-operative functional outcomes were worse. Microneurosurgical techniques and skills are critical to increase the total resection rate of posterior petrous meningiomas in order to decrease the mortality and disability rates.

Detection of cancer stem cells from the C6 glioma cell line.

Various malignant cancers have been found to contain a sub-population of stem cell-like tumour cells, or cancer stem cells (CSCs), however, culture methods for CSCs and the size of the fraction of CSCs in C6, which is a commonly used glioma cell line, remain controversial. In this study, we demonstrated that the C6 cell line contains a fraction of tumour cells that can form tumour spheres in a simplified serum-free neural stem cell medium and express CD133 and nestin, which are widely-used markers for brain CSCs. Immunohistochemistry and immunofluorescence confirmed the existence of CSCs both in the C6 cell line and C6 xenografts. Flow cytometry demonstrated that 4.02% of cells in the C6 cell line and 4.21% in the C6 xenografts presented as CSCs. These results confirm the fraction of CSCs in the C6 cell line and provide a simple and effective method for isolation of CSCs to study the initiation and progression of human glioma and, possibly, other malignant tumours.

Construction of tissue-engineered cartilage with BMP7 gene-transfected chondrocytes.

In this study, the expression of bone morphogenetic protein-7 (BMP7) was investigated in tissue-engineered cartilage constructed using chondrocytes transfected with the BMP7 gene and that constructed using non-transfected chondrocytes after 7, 14, 21 and 28 days. The volume of the BMP7 gene-transfected tissue-engineered cartilage culture after 5 - 7 days was 9 x 9 x 2 mm, while that of the non-transfected tissue-engineered cartilage culture was 8 x 8 x 2 mm. Histomorphological analysis showed that both cultures comprised cartilaginous tissue. Both BMP7 mRNA and BMP7 protein were expressed in BMP7 gene-transfected cartilage culture grown in vitro for 7, 14, 21 or 28 days. The chondrocytes in the BMP7 gene-transfected cartilage culture were active, with increased mitochondria, Golgi bodies and rough endoplasmic reticulum compared with non-transfected cartilage. These results provide an ideal foundation for the study of BMP7 gene-transfected tissue-engineered cartilage transplantation in the repair of cartilaginous defects.

[Synaptic connections between the neurons and catecholaminegic fibres in the hippocampal transplant of rat].

The foetal hippocampal tissue (transplant) at 17th day of embryonic age was implanted into the ventral hippocampus of adult rats (host). The catecholaminergic fibre projections in the hippocampal transplants 90 days after operation were studied by immunohistochemical technique. It was observed that in the host hippocampus there was a large population of TH- immunoreactive slender fibres of only 0.5-1 microns in diameter. These fibres were distributed more densely in the hippocampal hilus and CA3 transparent layers than in molecular layers, but sparsely in pyramidal or granular layers. In the molecular and cellular layers of transplanted hippocampus some thicker (> 1 micron) TH-positive fibers were ended in a relatively dense branching. The immunoelectron-microscopic observations showed that many TH-positive boutons made synaptic contacts with immunonegative dendrites and dendritic spines in the hippocampal transplants, the majority of which were asymmetrical synapses with a 30 nm synaptic cleft and conspicuously thickened postsynaptic membranes. It is concluded that catecholaminergic fibres extend from the host brain into the hippocampal transplant to establish synapses with the target neurons.

Quantitative investigation of the neuromuscular junction of rat skeletal muscle fibres after double innervation.

In normal (untreated) rats the mean length ratio of postsynaptic to presynaptic membrane was 2.7 +/- 0.8 for neuromuscular junctions of slow-twitch soleus muscle fibres and 4.2 +/- 1.0 for neuromuscular junctions of fast-twitch extensor digitorum longus muscle fibres; this difference was significant (P less than 0.001). After experimental double innervation by fast and slow muscle nerves for four months, the ratio was (1) 2.9 +/- 0.8 for the original slow-twitch fibre end-plate and 2.8 +/- 0.8 for the newly established one, both not significantly different from that of the normal slow-twitch fibres; and (2) 2.2 +/- 0.5 for the original fast-twitch fibre end-plate and 2.2 +/- 0.7 for the newly established one, both significantly smaller than that of the normal fast-twitch fibres (P less than 0.001). This means that the double innervated slow-twitch muscle fibres retained their original neuromuscular junction type, whereas the doubly-innervated fast-twitch muscle fibres underwent a dramatic transformation of their neuromuscular junction from the fast-muscle to the slow-muscle type. In both doubly innervated fibres, the ultrastructural characteristics of neuromuscular junctions, whether altered or not, were identical at both end-plate regions.