chondroitin-sulfates and Cadaver

McCarey-Kaufman's (MK) medium and Optisol-GS medium are the most commonly employed media for human donor corneal preservation. In this study, we evaluated the preservation efficacy of discarded human donor corneas using a Thermo-reversible gelation polymer (TGP) added to these two media.. Thirteen human corneal buttons collected from deceased donors, which were otherwise discarded due to low endothelial cell density (ECD) were used. They were stored in four groups: MK medium, MK medium with TGP, Optisol-GS and Optisol-GS with TGP at 4 °C for 96 h. Slit lamp examination and specular microscopy were performed. Corneal limbal tissues from these corneas were then cultured using explant methodology one with and the other without TGP scaffold, for 21 days.. MK + TGP and Optisol-GS + TGP preserved corneas better than without TGP, which was observed by maintenance of ECD which was significantly higher in Optisol-GS + TGP than MK + TGP (p-value = 0.000478) and corneal thickness remaining the same for 96 h. Viable corneal epithelial cells could be grown from the corneas stored only in MK + TGP and Optisol-GS + TGP. During culture, the TGP scaffold helped maintain the native epithelial phenotype and progenitor/stem cell growth was confirmed by RT-PCR characterization.. TGP reconstituted with MK and Optisol-GS media yields better preservation of human corneal buttons in terms of relatively higher ECD maintenance and better in vitro culture outcome of corneal limbal tissue. This method has the potential to become a standard donor corneal transportation-preservation methodology and it can also be extended to other tissue or organ transportation upon further validation.

Topics: Adult; Aged; Aged, 80 and over; Cadaver; Chondroitin Sulfates; Complex Mixtures; Culture Media; Dextrans; Endothelium, Corneal; Female; Gentamicins; Humans; Male; Middle Aged; Organic Chemicals; Slit Lamp Microscopy; Tissue Preservation

Ideally, cartilage regenerative cell therapy should produce a tissue which closely matches the microstructure of native cartilage. Benchmark reference information is necessary to assess the quality of engineered cartilage. Our goal was to examine the variation in glycosaminoglycans (GAGs) in cartilage zones within human knee joints of different ages.. Osteochondral biopsies were removed from the medial femoral condyles of deceased persons aged 20-50 years. Fluorophore-Assisted Carbohydrate Electrophoresis (FACE) was used to profile GAGs through the superficial, middle and deep zones of the articular cartilage. Differences were identified by statistical analysis.. Cartilage from the younger biopsies had 4-fold more hyaluronan in the middle zone than cartilage from the older biopsies. The proportion of hyaluronan decreased with increasing age. Cartilage from the middle and deep zones of younger biopsies had significantly more chondroitin sulphate and keratan sulphate than the cartilage from older biopsies. This would suggest that chondrocytes synthesise more sulphated GAGs when deeper in the tissue and therefore in conditions of hypoxia. With increasing age, there was significantly more chondroitin-6 sulphate than chondroitin-4 sulphate. For the first time, unsulphated chondroitin was detected in the superficial zone.. As an outcome measure, FACE offers the potential of a complete, detailed assessment of all GAGs and offers more information that the widely used 1,9-dimethylmethylene blue (DMMB) dye assay. FACE could be very useful in the evolving cartilage regeneration field.

Topics: Adult; Age Factors; Cadaver; Cartilage, Articular; Cell- and Tissue-Based Therapy; Chondroitin; Chondroitin Sulfates; Electrophoresis; Glycosaminoglycans; Guided Tissue Regeneration; Humans; Hyaluronic Acid; Keratan Sulfate; Knee Joint; Middle Aged; Outcome Assessment, Health Care; Reference Values; Tissue Engineering; Young Adult

This study was performed to evaluate the potential of a chondroitin sulfate-polyethylene glycol (CS-PEG) adhesive and collagen-based membrane (collagen vitrigel, CV) combination as a method to treat penetrating ocular injuries on the battlefield and to improve this method with two technologies: an antibiotic releasing CS-PEG adhesive and a corneal shaped CV. Burst testing using porcine cadaveric eyes, high-performance liquid chromatography, the Kirby-Bauer bacterial inhibition test, and CV implantations on the live and cadaveric rabbit eyes were performed. The ocular burst test showed CS-PEG adhesive could successfully repair 5-mm to 6-mm length wounds in the corneal and corneoscleral regions but would require CS-PEG + CV to treat larger wounds similar to those seen on the battlefield. In addition, high performance liquid chromatography and the Kirby-Bauer bacterial inhibition test presented evidence suggesting the vancomycin incorporated CS-PEG could inhibit Staphylococcus infection for 9 days. Furthermore, the curved CV showed an advantage by matching the corneal contour without any wrinkle formation. Although this pilot study showed a limited range of possible applications, we demonstrated that the combination of CS-PEG adhesive + CV is a promising method and the 2 technologies improve their applicability to the special demands of the battlefield.

Topics: Animals; Anti-Bacterial Agents; Blast Injuries; Cadaver; Chondroitin Sulfates; Collagen; Corneal Perforation; Disk Diffusion Antimicrobial Tests; Eye Injuries, Penetrating; Hydrogels; Male; Membranes, Artificial; Polyethylene Glycols; Rabbits; Staphylococcus aureus; Swine; Tissue Adhesives; Vancomycin

The most common site of rupture of the posterior tibial tendon is the retromalleolar region where the tendon changes its direction of pull. The aim of this study was to characterize the tissue of the gliding zone of the tibialis posterior tendon to gain further knowledge about possible structural causes for spontaneous tendon rupture.. Light microscopy, transmission electron microscopy and immunohistochemical methods were used to describe the structure of the human tibialis posterior tendon.. In the region where the tendon wraps around the medial malleolus, the structure of the tissue changes from the typical structure of a traction tendon. The superficial zone which was directed towards the pulley tissue had the structure of fibrocartilage with a specific three-dimensional collagen fibril texture. Transmission electron microscopy showed chondrocytes with a felt-like pericellular matrix that increased in size towards the gliding surface. The extracellular matrix of the fibrocartilage was rich in acid glycosaminoglycans and stained intensively with alcian blue at pH 1. Immunohistochemical staining of cartilage-specific extracellular matrix components such as type II collagen, chondroitin-4-sulphate, chondroitin-6-sulphate, keratan sulphate and aggrecan was positive.. The location of the fibrocartilage corresponds to the region where the tibialis posterior tendon wraps around the medial malleolus, which serves as a pulley. According to the theory of 'causal histogenesis', the stimulus for the development of fibrocartilage within dense connective tissue is intermittent compressive and shear stress. The fibrocartilaginous region is the region where most spontaneous ruptures of the tibialis posterior tendon occur. Due to its structure, the fibrocartilaginous region may be more vulnerable to repetitive tensile microtrauma; degeneration may occur due to the poor repair response of the avascular fibrocartilaginous tissue.

Topics: Adult; Aged; Aggrecans; Ankle Joint; Cadaver; Cartilage, Articular; Chondrocytes; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Extracellular Matrix; Extracellular Matrix Proteins; Female; Fibrillar Collagens; Glycosaminoglycans; Humans; Immunohistochemistry; Keratan Sulfate; Lectins, C-Type; Male; Microscopy; Middle Aged; Proteoglycans; Tendons

To compare the acute effects of Healon (sodium hyaluronate) and Viscoat (sodium chondroitin sulfate-sodium hyaluronate) on outflow facility in human cadaver eyes and determine which viscoelastic agent is least likely to cause an intraocular pressure (IOP) spike after cataract surgery.. The Glaucoma Research Lab, University of Toronto, Ontario, Canada.. In this prospective paired study, 15 pairs of human cadaver eyes were used. Following the construction of a 3.0 mm scleral tunnel, 0.25 cc of Healon was injected into the anterior chamber of 1 eye and 0.25 cc of Viscoat was injected into the contralateral eye. The viscoelastic agents were removed from both eyes in a standardized fashion and the scleral tunnels closed. The eyes were then perfused at a constant IOP of 8.0 mm Hg, corresponding to 16.0 mm Hg in vivo. Outflow facility (microL/minute [min]/mm Hg) was recorded every 15 minutes for 24 hours using standard methods.. Outflow facility in the Viscoat-treated eyes decreased appreciably for the first 3 hours, then recovered somewhat after 12 hours; facility in the Healon-treated eyes showed less of an overall decrease. Over the 24 hour perfusion period, mean outflow facility was 0.037 microL/min/mm Hg +/- 0.015 (SD) in the Viscoat-treated eyes and 0.060 +/- 0.012 microL/min/mm Hg in the Healon-treated eyes. Healon reduced outflow facility significantly less than Viscoat between 3.25 and 10.50 hours postoperatively (P < .05, 2-tailed t test).. Healon reduced outflow facility less than Viscoat between 3.25 and 10.50 hours postoperatively.

Topics: Anterior Chamber; Aqueous Humor; Cadaver; Chondroitin; Chondroitin Sulfates; Drug Combinations; Female; Humans; Hyaluronic Acid; In Vitro Techniques; Injections; Intraocular Pressure; Male; Prospective Studies; Time Factors; Trabecular Meshwork

Previous studies have presented evidence that an underlying cause of intervertebral disc degeneration is related to changes in the sulfation of the proteoglycans. The sulfation of the chondroitin in cadaveric lumbar intervertebral discs, at two different stages of degeneration as assessed by discogram, were analyzed. Fourteen of 28 lumbar discs were graded 2 and the other 14 were graded 4 (i.e., more degenerated). From each disc, six regional segments were carefully isolated. Proteoglycans were solubilized from the disc tissue with 4 M GuHCl. Chondroitin sulfate chains were analyzed by diethylaminoethyl (DEAE)-Sephacel and high-performance liquid chromatography (HPLC) anion exchange chromatography. The major differences in sulfation of the chondroitin between grade 2 and grade 4 discs only occurred in the posterior central annulus and nucleus segments. The chondroitin in the posterior central nucleus segments of the grade 2 and grade 4 intervertebral discs were undersulfated as compared with the other segments, and the differences between these segments and the others were more accentuated in the grade 4 discs than in grade 2 discs.

Topics: Cadaver; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Humans; Intervertebral Disc; Lumbar Vertebrae; Middle Aged; Radiography; Spinal Diseases

K-Sol corneal storage medium maintains corneal viability for at least ten days, compared with McCarey-Kaufman (M-K) medium, which does so for a reported maximum of three days. Since many eye banks still use M-K medium exclusively, stored corneas must be used without undue delay, and shipping time further hastens the planned surgery time. Transferring the shipped M-K medium-stored cornea to K-Sol offers a simple way of increasing storage time in our eye bank. Ninety-five out of 100 transferred corneas (95%) remained clear 3 to 18 months postoperatively. There were no primary graft failures, and transferred corneal mates were clear. A comparable series of 100 corneas stored in M-K medium alone, or in K-Sol alone, were clear in 93% and 97% of the transplants. Analysis of donor rims after keratoplasty indicated statistically significantly more endothelial cell loss during storage in corneas transferred to K-Sol compared with 18 corneas preserved only in M-K medium for a similar length of time. There was no statistical significance between the transferred group and the group preserved in only K-Sol. Transferring M-K medium stored corneas to K-Sol medium may be a useful means of extending corneal storage time, resulting in improved tissue utilization.

Topics: Cadaver; Chondroitin; Chondroitin Sulfates; Corneal Transplantation; Eye Banks; Graft Survival; Humans; Retrospective Studies; Tissue Preservation; Tissue Survival

The human intervertebral discs which were obtained by cadavers and anterior discectomy are investigated histochemically. Chondroitin-4S, chondroitin-6S, dermatan sulfate, hyaluronic acid and keratan sulfate were detected in the human intervertebral disc by various histochemical methods. pH2.5, pH1.1 toluidin blue metachromasia and 0.4M MgCl2 alcianophilia became weaker with increasing age, and the herniated disc were weaker than controlled discs in the same age group. Chondroitin-4S and chondroitin-6S were distributed throughout the discs. There was no clear localization of the various glycosaminoglycans in the human intervertebral disc, with the exception of keratan sulfate. There was no histologically and histochemically observable difference in the cervical and lumbar discs.

Topics: Adult; Cadaver; Cartilage; Chondroitin Sulfates; Dermatan Sulfate; Histocytochemistry; Humans; Hyaluronic Acid; Intervertebral Disc; Intervertebral Disc Displacement; Keratan Sulfate