Cell proliferation assays are essential to developing an understanding of the molecular mechanisms that modulate cell growth and differentiation. In this paper, we describe the application of alamarBlue, a new and versatile metabolic dye, for the detection of Swiss 3T3 fibroblast proliferation and/or survival. As a redox indicator, alamarBlue is reduced by reactions innate to cellular metabolism and, therefore, provides an indirect measure of viable cell number. Various assay parameters were optimized for a 96-well format to achieve a detectable range of fibroblast cell number from 100 to 20,000 cells/well, which is similar to that obtained with traditional (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and [3H]thymidine assay techniques. Standard (reference) curves generated with a known fibroblast stimulator were used to facilitate quantitation and comparison of unknown test substances. The alamarBlue assay offers the advantages of technical simplicity, freedom from radioisotopes, versatility in detection, no extraction, and excellent reproducibility and sensitivity. We anticipate that this simple and versatile alamarBlue assay, when used alone or in conjunction with other bioassays, will be a useful tool for investigating the complex mechanisms of cellular proliferation.

The importance of understanding cell-extracellular matrix (ECM) interaction is now evident as scientists, engineers, and physicians search for novel scaffolds that support and maintain tissue-specific cellular growth and function both in vivo and in vitro. Small intestinal submucosa (SIS) represents an ECM that has been derived from porcine intestine while preserving its natural composition and architecture. More recently, an extract of this physiologic ECM, which forms a three-dimensional gel in vitro, has been developed. When compared to routinely used culture substrata (e.g., plastic, Vitrogen, and Matrigel), intact SIS and SIS-derived gel possess unique compositional and architectural features. Simple squamous epithelial (pulmonary artery), fibroblastic (Swiss 3T3), glandular epithelial (adenocarcinoma), and smooth muscle-like (urinary bladder) cells were seeded upon intact SIS and SIS-derived gel and their morphologic response evaluated. For each of the four cell types studied, intact SIS and SIS-derived gel were equivalent or superior in their ability to support and maintain expression of tissue-specific phenotype when compared to the routinely used substrata, plastic, Vitrogen, and Matrigel. Therefore, SIS may provide a novel biologically derived scaffold for the growth and study of a variety of cell types in vitro. Such information regarding the influence of substrate structure and function on cell behavior will be useful in the development of successful tissue engineering strategies.

Osteochondral fragments were created arthroscopically on the distal aspect of both radial carpal bones in 12 horses. On day 14 after surgery, one middle carpal joint of each horse was injected with 2.5 mL Betavet Soluspan (3.9 mg betamethasone sodium phosphate and 12 mg betamethasone acetate per milliliter) and the contralateral joint was injected with 2.5 mL saline as a control. Intra-articular treatments were repeated on day 35. On day 17, six horses began exercising 5 days per week on a high-speed treadmill. The other six horses were kept in box stalls throughout the study as nonexercised controls. On day 56, all horses were examined clinically and radiographically and then were euthanatized. Samples were obtained for histological, histochemical, and biochemical evaluation. Mild lameness was observed in five of the six exercised horses at day 56; four horses were lame in the control limb and one horse was lame in the treated limb. Of the five nonexercised horses evaluated for lameness, two were lame in the control limb, two were lame in the treated limb, and one was lame in both the control and the treated limb. No differences were noted on radiographs or palpation of steroid treated limbs versus control limbs. Firm reattachment of the osteochondral fragment to the radial carpal bone occurred in all but three joints. Gross cartilage damage was not different between steroid-treated joints and joints injected with saline. Histologically, there were no significant detrimental effects of beta-methasone with or without exercise, but there was a tendency for more pathological change in treated joints. No significant difference in the water content or uronic acid concentration was detected between treated and control joints. Intra-articular betamethasone administration in this carpal chip model was not associated with any significant detrimental effects in either rested or exercised horses.

This study evaluated the use of computer-based interactive imagery on students' achievement scores when compared with paper-based static imagery. It also assessed students' perceptions about the two imagery strategies and their different components. Sixty-four freshmen veterinary students (50 females, 14 males), enrolled in a comparative anatomy course, volunteered to participate in the study. This study used a pretest/posttest comparison group design and data was examined by analysis of covariance (ANCOVA). A close-ended questionnaire was administered to collect students' perceptions about the two imagery strategies. The mean difference in students' perceptions between the two strategies was analyzed using a two-tailed paired t-test. No significant differences were observed between computer-based interactive imagery and paper-based static imagery in the immediate recall of anatomical information. There was a significant difference in students' opinions toward the two strategies: students perceived computer-based interactive imagery as a better strategy in the assimilation of anatomical information than paper-based static imagery.