Objective-To compare in vitro expansion of equine tendon- and bone marrow–derived cells with fibroblast growth factor-2 (FGF-2) supplementation and sequential matrix synthesis with pulverized tendon and insulin-like growth factor-I (IGF-I). Sample-Cells from 6 young adult horses. Procedures-Progenitor cells were expanded in monolayers with FGF-2, followed by culture with autogenous acellular pulverized tendon and IGF-I for 7 days. Initial cell isolation and subsequent monolayer proliferation were assessed. In pulverized tendon cultures, cell viability and expression of collagen types I and III and cartilage oligomeric matrix protein (COMP) mRNAs were assessed. Collagen and glycosaminoglycan syntheses were quantified over a 24-hour period. Results-Monolayer expansion with FGF-2 significantly increased the mean +/- SE number of tendon-derived cells (15.3 +/- 2.6 × 10(6)), compared with bone marrow-derived cells (5.8 +/- 1.8 × 10(6)). Overall, increases in collagen type III and COMP mRNAs were seen in tendon-derived cells, compared with results for bone marrow-derived cells. After IGF-I supplementation, increases in collagen type I and type III mRNA expression were seen in bone marrow–derived cells, compared with results for unsupplemented control cells. Insulin-like growth factor-I significantly increased collagen synthesis of bone marrow–derived cells. Monolayer expansion with FGF-2 followed by IGF-I supplementation significantly increased glycosaminoglycan synthesis in tendon-derived cells. Conclusions and Clinical Relevance-Tendon-derived cells had increased cell numbers and matrix synthesis after monolayer expansion with FGF-2, compared with results for bone marrow–derived cells. In vivo experiments with FGF-2-expanded tendon-derived cells are warranted to evaluate effects on tendon healing.

Objective: To assess the in vitro effects of various nalbuphine concentrations on viability and wound healing ability of corneal cells and potential drug transport through the corneal epithelium. Sample: Cultured canine and human corneal epithelial cells (CECs) and cultured canine corneal stromal fibroblasts. Procedures: CECs and stromal fibroblasts were exposed to nalbuphine (concentration of solutions ranged from 0% to 1.2%) for up to 30 minutes, and viability was assessed with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A standard scratch test technique was used. Wound healing of CECs and stromal fibroblasts was evaluated following treatment with nalbuphine solutions < 0.1%. Liquid chromatography–mass spectrometry–mass spectrometry analysis was used to evaluate drug transport across a monolayer and a multilayer of human CECs. Results: A progressive decrease in viability was detected in canine CECs for all nalbuphine treatment groups, whereas treatment with only 0.5% or 1.2% nalbuphine significantly reduced corneal stromal fibroblast viability, compared with results for control cells. Within 24 hours, treatment with 0.1% nalbuphine solution significantly altered the healing rate of both canine CECs and stromal fibroblasts. Continuous increases in transport rates of nalbuphine were detected with time for both the monolayer and multilayer of human CECs. Conclusions and Clinical Relevance: In vitro, nalbuphine potentially could penetrate through corneal tissue, but it may cause damage to the corneal epithelium and stromal fibroblasts. Therefore, nalbuphine potentially may impair corneal wound healing.

Objective: To determine the distribution of cannabinoid receptor type 1 (CB1) and cannabinoid receptor type 2 (CB2) in skin (including hair follicles and sweat and sebaceous glands) of clinically normal dogs and dogs with atopic dermatitis (AD) and to compare results with those for positive control samples for CB1 (hippocampus) and CB2 (lymph nodes). Sample: Skin samples from 5 healthy dogs and 5 dogs with AD and popliteal lymph node and hippocampus samples from 5 cadavers of dogs. Procedures: CB1 and CB2 were immunohistochemically localized in formalin-fixed, paraffin-embedded sections of tissue samples. Results: In skin samples of healthy dogs, CB1 and CB2 immunoreactivity was detected in various types of cells in the epidermis and in cells in the dermis, including perivascular cells with mast cell morphology, fibroblasts, and endothelial cells. In skin samples of dogs with AD, CB1 and CB2 immunoreactivity was stronger than it was in skin samples of healthy dogs. In positive control tissue samples, CB1 immunoreactivity was detected in all areas of the hippocampus, and CB2 immunoreactivity was detected in B-cell zones of lymphoid follicles. Conclusions and Clinical Relevance: The endocannabinoid system and cannabimimetic compounds protect against effects of allergic inflammatory disorders in various species of mammals. Results of the present study contributed to knowledge of the endocannabinoid system and indicated this system may be a target for treatment of immune-mediated and inflammatory disorders such as allergic skin diseases in dogs.

A number of studies have measured collagen fibers and collagen&#xD;deposition in wound healing process with advances imaging techniques. However, these are performed by complicated methods and need specific tools. In search of the easier ways in routine histopathological laboratory, collagen measurement and staining pattern of wound healing process were observed in wounded skin of Sprague Dawley’s rat by using two different stains which are standard haematoxylin and eosin (H&E) and modified Masson’s&#xD;trichrome staining (MT). The comparison between these staining in wounded tissues was made to evaluate the advantages and disadvantages of both staining in wound healing study for 21 days postwounding. Tissues which stained with MT staining was then evaluated its collagen re-organization and density by using polarized light microscope with the aid of image analyzer software. Results showed that tissues stained with standard H&E could not be used to measure and differentiate the collagen deposition which is contradictory to MT staining. Wounded tissue stained with MT staining has showed a clear view of collagen fibers deposition&#xD;and re-organisation compared to H&E staining. This finding could validate the using of modified MT staining which leads to accurate histopathological analysis and observation in wound healing study.