Methodological models for in vitro amplification and maintenance of human articular chondrocytes from elderly patients.

Articular cartilage defects, an exceedingly common problem closely correlated with advancing age, is characterized by lack of spontaneous resolution because of the limited regenerative capacity of adult articular chondrocytes. Medical and surgical therapies yield unsatisfactory short-lasting results. Recently, cultured autologous chondrocytes have been proposed as a source to promote repair of deep cartilage defects. Despite encouraging preliminary results, this approach is not yet routinely applicable in clinical practice, but for young patients. One critical points is the isolation and ex vivo expansion of large enough number of differentiated articular chondrocytes. In general, human articular chondrocytes grown in monolayer cultures tend to undergo dedifferentiation. This reversible process produces morphological changes by which cells acquire fibroblast-like features, loosing typical functional characteristics, such as the ability to synthesize type II collagen. The aim of this study was to isolate human articular chondrocytes from elderly patients and to carefully characterize their morphological, proliferative, and differentiative features. Cells were morphologically analyzed by optic and transmission electron microscopy (TEM). Production of periodic acid-schiff (PAS)-positive cellular products and of type II collagen mRNA was monitored at different cellular passages. Typical chondrocytic characteristics were also studied in a suspension culture system with cells encapsulated in alginate-polylysine-alginate (APA) membranes. Results showed that human articular chondrocytes can be expanded in monolayers for several passages, and then microencapsulated, retaining their morphological and functional characteristics. The results obtained could contribute to optimize expansion and redifferentiation sequences for applying cartilage tissue engineering in the elderly patients.

Scanning electron microscopic analysis of acrylic intraocular lenses for microincision cataract surgery.

PURPOSE: To analyze the surface quality before and after folding of 2 intraocular lens (IOL) models designed for microincision cataract surgery. SETTING: Eye Clinic and Department of Human Pathology and Oncology, University of Florence, Florence, Italy. METHODS: Two IOL models now available for sub-2.0 mm microincision were studied: UltraChoice 1.0 Rollable ThinLens (ultrathin lens by ThinOptX) and AcriSmart (foldable lens by AcriTec). Eight IOLs of each model were examined. Scanning electron microscopy (SEM) was performed before and after IOL folding with a dedicated injector. Special attention was given to the optic surface and edges, the optic-haptic junction, and the haptic itself. RESULTS: Initially, the surface quality of IOLs was evaluated before folding. On SEM, smooth and homogeneous optic and haptic surfaces were revealed in both IOL models with no surplus material or molding flashes; edge finish of all optics showed no evidence of ridges. The IOL surfaces were evaluated after insertion into their injectors and after ejection at room temperature. After folding, the microincision IOLs showed no sign of surface alteration, probably because of their high water content, which makes these IOLs soft and flexible. CONCLUSIONS: Visual results and long-term biocompatibility of the IOLs are influenced by surface properties. In recent years, there has been a trend toward microincision cataract surgery. Our study shows that the 2 IOL models now available for sub-2.0 mm microincision have acceptable surface properties.

Folding procedure for acrylic intraocular lenses.

PURPOSE: To compare in vitro the effect of 2 standard methods of folding acrylic intraocular lenses (IOLs) on surface characteristics and bacterial adhesion. SETTING: Eye Clinic and Department of Health-Microbiology Unit, University of Florence, Florence, Italy. METHODS: To evaluate the effect of folding, 2 types of acrylic IOLs were not folded or folded with a forceps or an injector and then processed for scanning electron microscopy (SEM) examination. Bacterial adhesion was assessed using an ocular isolate of Pseudomonas aeruginosa. Nonfolded and folded IOLs were placed in test tubes containing the bacterial suspension for direct counting of viable adherent bacteria and for SEM. RESULTS: The injector-folded IOLs did not show major alterations on the surface; 5 of the 9 forceps-folded IOLs showed marks or scratches in the profile of the optic. The mean number of viable adherent bacteria per area of IOL optic was 1082 (95% confidence interval [CI], 835-1330) in forceps-folded IOLs, 366 (95% CI, 192-359) in injector-folded IOLs, and 206 (95% CI, 123-289) in nonfolded IOLs. Scanning electron microscopy confirmed more surface irregularities on forceps-folded IOLs, with bacteria adherent preferentially on the surface scratches. CONCLUSION: Forceps-folding provoked more surface irregularities, which probably make IOLs more susceptible to bacterial adhesion.