Patterns of Hyaluronan Staining Are Modified by Fixation Techniques

Lin, Wenming; Shuster, Svetlana; Maibach, Howard I.; Stern, Robert

doi:10.1177/002215549704500813

Cited by 124 publications

(105 citation statements)

References 20 publications

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“…HABP is a complex of aggrecan and link protein, and is derived from bovine cartilage. [35][36][37] In our study, using this specific probe, the surfaces of the tendons with more HA (Groups 0.50:0.25, 0.50:1.00, 1.00:0.25, 1.00:1.00, and 2.00:1.00) were smoother and had lower gliding resistance after 1000 cycles compared to the groups with less HA retained on the surface after 1000 cycles.…”

Section: Discussionmentioning

confidence: 59%

Optimization of surface modifications of extrasynovial tendon to improve its gliding ability in a canine model in vitro

et al. 2006

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Previous studies have shown that the carboxyl groups in hyaluronic acid (HA) could be activated by 1-ethy 1-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) to form intermediate O-acylisoureas, which can chemically bind to exposed amino groups on the tendon surface, leading to improved gliding ability. However, the optimal ratio and concentrations of the components in this chemical mixture were not investigated. The purpose of this study was to optimize the constituents of this tissue engineering approach to tendon surface modification, to reduce friction and improve durability. Peroneus longus (PL) tendons (n ¼ 40) were harvested from adult mongrel dogs along with the A2 pulley obtained from the ipsilateral hind paw. After the gliding resistance of the normal PL tendon was measured, the tendons were treated under varying concentrations of HA (0.5, 1, and 2%) and EDC/NHS (0.05, 0.25, and 1%) mixed with a 10% gelatin. Tendon friction was measured for 1000 cycles of simulated flexion/extension motion. Following testing, the residual HA on the tendon surface was evaluated by immunohistochemisty. The gliding resistance of the untreated PL tendons had a mean value of 0.087 AE 0.021 N. After surface treatment, there was no significant difference in friction due to HA concentration alone, but the concentration of EDC/NHS and the interaction between HA concentration and EDC/NHS concentration had a significant effect on friction. Regardless of HA concentration, the friction after 1000 cycles was significantly decreased in preparations which included a 1% concentration of EDC/ NHS. The tendons with lower gliding resistance presented a smoother surface on light microscopy and maintained more residual HA on the tendon surface. By varying the relative concentrations of HA, EDC, and NHS it is possible to optimize the effect of surface treatment on friction and durability in a canine extrasynovial tendon in vitro. ß

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Section: Discussionmentioning

confidence: 59%

Optimization of surface modifications of extrasynovial tendon to improve its gliding ability in a canine model in vitro

et al. 2006

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“…Spent media samples were assayed for glucose (Amplex Red Assay; Invitrogen), GAG, and OHP. Samples for histological characterization were fixed (5% acetic acid, 3.7% formaldehyde, 70% ethanol solution), 35 dehydrated in successively concentrated ethanol solutions, embedded in paraffin, and sectioned to 7 mm slices. Following paraffin removal, sections were stained with either Safranin O to qualitatively assay the negative charges of GAG or Picrosirius red to qualitatively assay for collagen.…”

Section: Mechanical Biochemical and Histological Measurementsmentioning

confidence: 99%

Matrix Production in Large Engineered Cartilage Constructs Is Enhanced by Nutrient Channels and Excess Media Supply

Nims

Cigáň

Albro

et al. 2015

Tissue Engineering Part C: Methods

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Cartilage tissue engineering is a promising approach to resurfacing osteoarthritic joints. Existing techniques successfully engineer small-sized constructs with native levels of extracellular matrix (glycosaminoglycans [GAG] or collagen). However, a remaining challenge is the growth of large-sized constructs with properties similar to those of small constructs, due to consumption and transport limitations resulting in inadequate nutrient availability within the interior of large constructs. This study employed system-specific computational models for estimating glucose requirements of large constructs, with or without channels, to enhance nutrient availability. Based on glucose requirements for matrix synthesis in cartilage constructs, computational simulations were performed to identify the media volume (MV) and the number of nutrient channels (CH) needed to maintain adequate glucose levels within tissue constructs over the 3-day period between media replenishments. In Study 1, the influence of MV (5, 10, 15 mL/construct) and number of nutrient channels (CH: 0, 3, 7, 12 per construct) on glucose availability was investigated computationally for B10 · 2.34 mm cylindrical constructs. Results showed that the conventionally used MV 5 led to deleterious glucose depletion after only 40 h of culture, and that MV 15 was required to maintain sufficient glucose levels for all channel configurations. Study 2 examined experimentally the validity of these predictions, for tissue constructs cultured for 56 days. Matrix elaboration was highest in MV 15/CH 12 constructs (21.6% -2.4%/ww GAG, 5.5% -0.7%/ww collagen, normalized to wet weight (ww) on day 0), leading to the greatest amount of swelling (3.0 -0.3 times day-0 volume), in contrast to the significantly lower matrix elaboration of conventional culture, MV 5/CH 0 (11.8% -1.6%/ww GAG and 2.5% -0.6%/ww collagen, 1.6 -0.1 times day-0 volume). The computational analyses correctly predicted the need to increase the conventional media levels threefold to support matrix synthesis in large channeled engineered constructs. Results also suggested that more elaborate computational models are needed for accurate predictive tissue engineering simulations, which account for a broader set of nutrients, cell proliferation, matrix synthesis, and swelling of the constructs.

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“…20 Staining was similar to that described previously. 21 Slides were incubated in 3% BSA for 30 min, rinsed with PBS-CMF and then 0.3 ml of the diluted biotinylated hyaluronan binding peptide solution (1:200 of a 0.5 mg/ml preparation) and incubated overnight at 4°C. Slides were then washed for 10 min with calcium-and magnesium-free PBS.…”

Section: Histochemistry Of Hyaluronanmentioning

confidence: 99%

Hyaluronidase reduces human breast cancer xenografts in SCID mice

Shuster

Frost

Csóka

et al. 2002

Intl Journal of Cancer

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A hyaluronan-rich environment often correlate with tumor progression. and may be one mechanism for the invasive behavior of malignancies. Eradication of hyaluronan by hyaluronidase administration could reduce tumor aggressiveness and would provide, therefore, a new anti-cancer strategy. Hyaluronan interaction with its CD44 receptor and the resulting signal transduction events may be among the mechanisms for hyaluronan-associated cancer progression. We have shown previously that hyaluronidase treatment of breast cancer cells in vitro not only eradicates hyaluronan but also modifies expression of CD44 variant exons of tumor cells. We now determine if such effects occur in vivo and if it is accompanied by tumor regression. SCID mice bearing xenografts of human breast carcinomas were given intravenous hyaluronidase. Tumor volumes decreased 50% in 4 days. Tumor sections showed decreased hyaluronan. Intensity of staining for CD44s was not affected, whereas staining for specific CD44 variant exon isoforms was greatly reduced in residual tumors. Necrosis was not evident. Hyaluronidase, used previously as an adjunct in cancer treatment, presumably to enhance penetration of chemotherapeutic drugs, may itself have intrinsic anti-cancer activity. Removing peritumor hyaluronan appears to cause an irreversible change in tumor metabolism. Continuous hyaluronan binding to CD44 variant exon isoforms may also be required to stabilize inherently unstable isoforms that participate perhaps in tumor progression. Further investigation is required to confirm a cause and effect relationship between loss of hyaluronan, changes in CD44 variant exon expression and tumor reduction. If confirmed, hyaluronidase may provide a new class of anti-cancer therapeutics and one without toxic side effects.

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Patterns of Hyaluronan Staining Are Modified by Fixation Techniques

Cited by 124 publications

References 20 publications

Optimization of surface modifications of extrasynovial tendon to improve its gliding ability in a canine model in vitro

Optimization of surface modifications of extrasynovial tendon to improve its gliding ability in a canine model in vitro

Matrix Production in Large Engineered Cartilage Constructs Is Enhanced by Nutrient Channels and Excess Media Supply

Hyaluronidase reduces human breast cancer xenografts in SCID mice

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