Physicochemical and structural analysis of three regions of the deep digital flexor tendon of pigs

Few studies have discussed the relationship between the molecular organization and the physicochemical and biomechanical properties of pig tendons. In this work, we examined the extracellular matrix of the deep digital flexor tendon of pigs, which was subjected to tensional (proximal region) and compressive (distal and terminal regions) forces. The three regions of the tendon were used for swelling tests and their glycosaminoglycan content was determined. Longitudinal sections of the tendon were stained and observed using polarized light microscopy. The distal and terminal regions were swole more in water than the proximal region. After staining with toluidine blue the metachromasy was more intense in the distal and terminal regions, indicating an accumulation of proteoglycans in these regions. Analysis of the glycosaminoglycans by agarose gel electrophoresis showed that dermatan sulfate was present in all regions, whereas chondroitin sulfate occurred only in the regions of compression. The shape of the fibroblasts changed along the tendon: rounded cells occurred in regions under compression, while in the region under tension, elongated cells predominated. The organization and distribution of the collagen bundles were different for each region. Birefringence analysis revealed a more regular crimp pattern in the region under tension than in the regions under compressive forces. The elastic fibers also showed a different distribution in each region. These results indicate that the regional differences in the structure and composition of the deep digital flexor tendon of pigs are related to the biomechanical properties of the tendon.

Extracellular matrix of ostrich articular cartilage

The composition and organization of the extracellular matrix of ostrich articular cartilage was investigated, using samples from the proximal and distal surfaces of the tarsometatarsus. For morphological analysis, sections were stained with toluidine blue and analyzed by polarized light microscopy. For biochemical analysis, extracellular matrix components were extracted with 4 M guanidinium chloride, fractionated on DEAE-Sephacel and analyzed by SDS-PAGE. Glycosaminoglycans were analyzed by electrophoresis in agarose gels. Structural analysis showed that the fibrils were arranged in different directions, especially on the distal surface. The protein and glycosaminoglycan contents of this region were higher than in the other regions.SDS-PAGE showed the presence of proteins with molecular masses ranging from 17 to 121 kDa and polydisperse components of 67, 80-100, and 250-300 kDa in all regions. The analysis of glycosaminoglycans in agarosepropylene diamine gels revealed the presence of only chondroitin-sulfate. The electrophoretic band corresponding to putative decorin was a small proteoglycan containing chondroitin-sufate and not dermatan-sulfate, unlike other cartilages. The higher amounts of proteins and glycosaminoglycans and the multidirectional arrangement of fibrils seen in the distal region may be correlated with the higher compression normally exerted on this region

Ultrastructural characteristics of tensional regions in tendons from rats of different ages

The calcaneal tendon and the deep digital flexor tendon are collagen-rich structures which are adapted to resist tensile stress. Since during aging tendons undergo modifications in their mechanical properties and in collagen aggregation, an understanding of the structural changes involved is important. In this work, the structural organization of the tensile region of the calcaneal and deep difital flexor tendons was studied in male Wistar rats 30, 180 and 730 days old. Large quantities of rough endoplasmic reticulum and peripheral secretory microvesicles were observed in the calcaneal tendon of 30-day-old rats. In the case of the deep digital flexor tendon, this organelle remained well-developed up to 180 days. A marked decrease in rough endoplasmic reticulum was observed in both tendons in 730-day-old rats. Proteoglycans associated with collagen fibrils were visible in the two tendons of all age groups. The reduced amount of rough endoplasmic reticulum and secretory microvesicles may be correlated with the known lower turnover of extracellular matrix components during aging.

Effect of magnesium chloride and guanidinum chloride on the extraction of components of extracellular matrix from chicken cartilage

In order to evaluate the effect of chaotropic agents on proteoglycan and non-collagenous proteins, chicken xiphoid cartilage was treated with guanidine-HCI and MgCl2 in different concentrations (1M to 5M), and different periods of time (12, 24, 48 and 72hr). The maximum yield of uronic acid was obtained with 3M MgCl2 (73.3 per cent). Concentrations of 4M and 5M of MgCl2 showed that much less uronic acid was removed, 55.3 per cent and 38.1 respectively. Extraction with 3M MgCl2 and 3M guanidine-HCl resulted better efficiency when performed for 48 hr. Analysis by SDS-PAGE of the extracts obtained with guanidine-HCl and MgCl, in different concentrations pointed out that most components are equally removed with the two solvents, showing that the extraction with MgCl2 is an alternative assay to remove non-collagenous proteins from extracellular matrix