Papers by Author: Traian V. Chirila

Abstract: A series of linear poly(2-hydroxyethyl methacrylate) (PHEMA) with defined molecular weights (MW) and narrow molecular distributions were synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization using cumyl dithiobenzoate (CDB) as a chain transfer agent. Murine fibroblasts (3T3) were exposed to eluates from various PHEMA samples, washed or unwashed, and with or without dithioester end groups. After 72 hrs in cell culture, no cytotoxic response was elicited by the polymer samples devoid of dithioester end groups, and which also underwent a thorough washing regime. Specimens throughout the entire MW range were internalized by a macrophage (cell line Raw 264), suggesting that such polymers can be used as models for studying the biodegradation of PHEMA.

Abstract: A nonapeptide, which is sensitive to enzymatic digestion by collagenase, was modified by the covalent attachment of an acrylamido group at the terminal positions. The functionalized peptide was used as a crosslinking agent during polymerization of 2-hydroxyethyl methacrylate (HEMA). Reversible addition-fragmentation chain transfer (RAFT) method was used to obtain a polymer (PHEMA) with an average theoretical molecular weight of 4000 Da, containing enzymatically labile peptide crosslinks. The functionalized peptide was analyzed in detail by 1H and 13C nuclear magnetic resonance (NMR) spectrometry. The polymerization reaction was monitored by near infrared spectrometry, while the resulting polymer was analyzed by size exclusion chromatography and solid NMR spectrometry. The peptide-crosslinked PHEMA was subjected to an in-vitro degradation assay in the presence of collagenase. At the highest concentration of enzyme used in the study, a weight loss of 35% was recorded after 60 days of incubation in the collagenolytic medium. This suggests that crosslinking with enzymatically degradable peptides is a valid method for inducing biodegradability in polymers that otherwise are not degradable.

Abstract: Dysfunction of the corneal endothelium due to cell loss caused by aging, disease or trauma can lead to severe visual impairment and blindness. Traditionally, dysfunctional endothelia are managed surgically, by removing the entire central cornea and transplanting either donor corneal tissue (penetrating keratoplasty), or just endothelia isolated from donor corneas. As in many cases it is only the corneal endothelium requiring replacement, many attempts were made over the last decades to develop an endothelial substitute, thereby precluding the need for the use of full donor corneas. This article reviews these attempts, which include artificial membranes, cell-coated corneal transplants, and cell-coated membranes. The presumption of an artificial corneal endothelium capable of duplicating the transendothelial ion-and-fluid transport function is examined in light of the latest hypotheses regarding the mechanism of this function.

Abstract: Silk fibroin (SF), isolated from silkworm (Bombyx mori) cocoons, is a natural biodegradable polymer. Over the past decade, there was some interest in using SF as a biomedical material. As part of a project to develop tissue-engineered constructs for the surgical restoration of the ocular surface (cornea, conjunctiva), we have investigated the capacity of SF to function as a substratum for the attachment and growth of corneal stem/progenitor cells harvested from the corneoscleral limbus of donor human corneal tissue. SF membranes were produced from cocoons following a protocol involving successive dissolution steps, filtration, dialysis, evaporation, and methanol treatment. Human limbal epithelial cells were harvested from donor tissue and seeded onto SF membranes. After 5 days, the culture was fixed and stained with specific agents to visualize the cells. The study indicated profuse cellular attachment and growth. SF membranes appear to be suitable as a substratum for the repair of damaged ocular surface.