No abstract

The in situ-gelling chitosan solution described here can support in vitro and in vivo accumulation of cartilage matrix by primary chondrocytes, while persisting in osteochondral defects at least 1 week in vivo.

Solidification of a chitosan-glycerol phosphate/blood implant in microfracture defects improved cartilage repair compared with microfracture alone by increasing the amount of tissue and improving its biochemical composition and cellular organization.

Chitosan-GP/blood implants applied in conjunction with drilling, compared to drilling alone, elicited a more hyaline and integrated repair tissue associated with a porous subchondral bone replete with blood vessels. Concomitant regeneration of a vascularized bone plate during cartilage repair could provide progenitors, anabolic factors and nutrients that aid in the formation of hyaline cartilage.

The MMTVD/myc transgenic mice spontaneously develop oligoclonal CD4+CD8 + T-ceU tumors. We used provirus insertional mutagenesis in these mice to identify putative collaborators of c-myc. We found that Notchl was mutated in a high proportion (52%) of these tumors. Proviruses were inserted upstream of the exon coding for the transmembrane domain and in both transcriptional orientations. These mutations led to high expression of truncated Notchl RNAs and proteins (86-110 kD). In addition, many Notchl-rearranged tumors showed elevated levels of full-length Notchl transcripts, whereas nearly all showed increased levels of full-length (330-kD) or close to full-length (280-kD) Notchl proteins. The 5' end of the truncated RNAs were determined for some tumors by use of RT-PCR and 5' RACE techniques. Depending on the orientation of the proviruses, viral LTR or cryptic promoters appeared to be utilized, and coding potential began in most cases in the transmembrane domain. Pulse-chase experiments revealed that the 330-kD Notchl proteins were processed into 110-and 280-kD cleavage products. These results suggest that Notchl can be a frequent collaborator of c-myc for oncogenesis. Furthermore, our data indicate that Notchl alleles mutated by provirus insertion can lead to increased expression of truncated and full-length (330/280-kD) Notchl proteins, both being produced in a cleaved and uncleaved form.

Bone marrow stimulation is performed using several surgical techniques that have not been systematically compared or optimized for a desired cartilage repair outcome. In this study, we investigated acute osteochondral characteristics following microfracture and comparing to drilling in a mature rabbit model of cartilage repair. Microfracture holes were made to a depth of 2 mm and drill holes to either 2 mm or 6 mm under cooled irrigation. Animals were sacrificed 1 day postoperatively and subchondral bone assessed by histology and micro-CT. We confirmed one hypothesis that microfracture produces fractured and compacted bone around holes, essentially sealing them off from viable bone marrow and potentially impeding repair. In contrast, drilling cleanly removed bone from the holes to provide access channels to marrow stroma. Our second hypothesis that drilling would cause greater osteocyte death than microfracture due to heat necrosis was not substantiated, because more empty osteocyte lacunae were associated with microfracture than drilling, probably due to shearing and crushing of adjacent bone. Drilling deeper to 6 mm versus 2 mm penetrated the epiphyseal scar in this model and led to greater subchondral hematoma. Our study revealed distinct differences between microfracture and drilling for acute subchondral bone structure and osteocyte necrosis. Additional ongoing studies suggest these differences significantly affect long-term cartilage repair outcome. ß

No abstract

Meniscus injury is a frequently encountered clinical orthopedic issue and is epidemiologically correlated to osteoarthritis. The development of new treatments for meniscus injury is intimately related to the appropriateness of animal models for their investigation. The purpose of this study was to structurally compare human menisci to sheep and rabbit menisci to generate pertinent animal models for meniscus repair. Menisci were analyzed histologically, immunohistochemically, and by environmental scanning electron microscopy (ESEM). In all species, collagen I appeared throughout most menisci, but was absent from the inner portion of the tip in some samples. Collagen II was present throughout the inner main meniscal body, while collagen VI was found in pericellular and perivascular regions. The glycosaminoglycan-rich inner portion of menisci was greater in area for rabbit and sheep compared to human. Cells were rounded in central regions and more fusiform at the surface, with rabbit being more cellular than sheep and human. Vascular penetration in rabbit was confined to the very outermost region (1% of meniscus length), while vessels penetrated deeper into sheep and human menisci (11-15%). ESEM revealed a lamellar collagenous structure at the articulating surfaces of sheep and human menisci that was absent in rabbit. Taken together, these data suggest that the main structural features that will influence meniscus repair-cellularity, vascularity, collagen structure-are similar in sheep and human but significantly different in rabbit, motivating the development of ovine meniscus repair models. ß