Factors affecting rotator cuff healing.

Several studies have noted that increasing age is a significant factor for diminished rotator cuff healing, while biomechanical studies have suggested the reason for this may be an inferior healing environment in older patients. Larger tears and fatty infiltration or atrophy negatively affect rotator cuff healing. Arthroscopic rotator cuff repair, double-row repairs, performing a concomitant acromioplasty, and the use of platelet-rich plasma (PRP) do not demonstrate an improvement in structural healing over mini-open rotator cuff repairs, single-row repairs, not performing an acromioplasty, or not using PRP. There is conflicting evidence to support postoperative rehabilitation protocols using early motion over immobilization following rotator cuff repair.

Sensory response following knee joint damage in rabbits.

BACKGROUND: Altered sensory information arising from damaged knee joint structures has been hypothesized as a contributing factor to persistent muscle dysfunction following injury. METHODS: Composite femoral nerve sensory signal was measured in 24 rabbits randomly allocated (8 per group) to receive surgical anterior cruciate ligament (ACL) transection with or without autograft reconstruction or nothing (control). Two-weeks after the intervention composite afferent signals were recorded from the femoral nerve. Side-to-side ratios (surgical side vs contralateral healthy side) for peak femoral nerve afferent composite signal were used for comparison. RESULTS: Femoral nerve afferent signal ratios were significantly higher in the ACL-R (2.21 ± 0.74) group when compared to the ACL-T (1.28 ± 0.61, P=0.02) group and Control group (1.31 ± 0.78, P=0.03). CONCLUSION: The magnitude of sensory information recorded on the femoral nerve is increased following ACL injury and reconstruction surgery, but not after an isolated ACL injury in rabbits.

Arthroscopic rotator cuff repair: suture anchor properties, modes of failure and technical considerations.

Rotator cuff injury and tears are a common source of shoulder pain, particularly among the elderly. Arthroscopic repair has now become the mainstay in the treatment of significant injuries that have failed conservative therapy. Compared with the traditional open technique, arthroscopic repair offers patients smaller incisions and less soft-tissue trauma, which result in improved postoperative pain and rehabilitation. The advances that have made arthroscopic repairs a reality includes improvement in arthroscopic rotator cuff instrumentation, particularly suture anchors. Suture anchors are used to reattach the torn rotator cuff tissue back onto the bone. Current rotator cuff anchors vary by design, anchor composition and suture materials. A treating physician should be aware of the advantages and limitations of these implants, which may influence the choice of one anchor over another. In addition to anchor variables, other factors that may affect the success of the repair include the local environment and surgical technique. In this article, various aspects of anchor design will be discussed. In addition, a concise review of technical considerations will also be discussed.

Recombinant growth/differentiation factor-5 stimulates osteogenic differentiation of fat-derived stromal cells in vitro.

Fat-derived stromal cells can differentiate into various skeletal tissues. Currently the mechanism that determines whether stromal cells differentiate into osteoblasts is unclear and the role of growth/differentiation factor (GDF)-5 in differentiation of fat-derived stromal cells is not fully understood. It appears that the differentiation of stromal cells is greatly enhanced by GDF-5 that plays a role in a variety of musculoskeletal processes such as joint formation, tendon maintenance, and bone formation. Our study showed that GDF-5 promotes the differentiation of rat fat-derived stromal cells into osteogenic lineages in vitro. Furthermore, these findings were confirmed by histology, biochemical assay for alkaline phosphatase activity, and analysis of gene expression. The ability to preferentially stimulate fat-derived stromal cells down the osteogenic pathway holds significance in a variety of clinical scenarios.

Osteogenic differentiation of adipose-derived stromal cells treated with GDF-5 cultured on a novel three-dimensional sintered microsphere matrix.

BACKGROUND CONTEXT: It is well known that under the proper conditions multipotential bone marrow stromal cells are capable of osteogenic differentiation. Recently studies have demonstrated that an analogous subpopulation of cells exist within adipose tissue. Although early studies characterizing these adipose-derived stromal (ADS) cells in culture exist, investigations exploring the characteristics and viability of these cells cultured on a three-dimensional sintered microsphere matrix are absent. PURPOSE: To characterize and investigate the viability of ADS cells cultured on bioengineered three-dimensional sintered microsphere matrices (SMM). STUDY DESIGN: Basic science, laboratory study. PATIENT SAMPLE: Sixty SMM total. Six underwent examination by scanning electron microscopy, 18 for cellular viability, 18 for biochemical assay, and 18 for evaluation by gene expression. OUTCOME MEASURES: The SMM were examined under scanning electron microscopy to evaluate for adherence, migration, and proliferation at 7, 14, and 28 days. Cellular viability was assessed using colorimetric assay for mitochondrial dehydrogenases activity in viable cells (MTS [3-(4,5-dimethylthiazol-2-yl)5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay) at each corresponding time point. Osteoblastic differentiation was determined using biochemical assays for alkaline phosphatase activity and gene expression for alkaline phosphatase (ALP), osteocalcin (OC), and core binding factor alpha-1 (Cbfa1). METHODS: Multipotential ADS cells from adult Sprague Dawley rats were isolated and maintained in media. Sintered microsphere matrices of poly(lactide-co-glycolide) [85:15] were prepared using solvent evaporation technique followed by mechanical sieving and fabricated by heating in metal molds. ADS cells were then seeded on the SMM and cultured in media with growth and differentiation factor-5 (GDF-5). Treated samples and controls were evaluated at 7, 14, and 28 days. Statistical significance was set at p<.05. RESULTS: Multipotential ADS cells were capable of being isolated from adipose tissue. Scanning electron microscopy evaluation revealed cells adherent to the scaffold surface in a monolayer by 7 days. Cytoplasmic extensions were seen linking the cells on adjacent microspheres. Migration and proliferation resulting in extension of the cellular elements into the scaffold was apparent by 14 days. MTS confirmed cell viability within the scaffold throughout the 28-day study. Osteoblastic differentiation was confirmed using biochemical assays for alkaline phosphatase activity and gene expression for ALP, OC, and Cbfa1. CONCLUSIONS: This is the first study to investigate the fate of ADS seeded on a three-dimensional sintered microsphere matrix. The results of this study confirm that ADS cells, when treated with GDF-5, are not only capable of adhering to the bioengineered scaffold, but also remain viable and demonstrated the ability to migrate, proliferate, and subsequently undergo osteogenic differentiation under the conditions described. These early findings support the concept that ADS cells cultured on a SMM may serve as a viable alternative to more traditional methods of bone graft materials.

Treatment options in pediatric femoral shaft fractures.

Fracture of the femur in a pediatric patient presents special problems, and a variety of treatment options. Child abuse and neglect should be considered and evaluated. Fractures in infants (0-18 months) may be treated successfully in a Pavlik Harness. Spica casting is safe and effective in children up to about 6 years or 100 pounds, although complications can occur and careful attention to technique is important. Surgical treatment is superior in most older or larger children or adolescents, and in cases of multiple trauma, soft tissue injury, obesity or head injury. External fixation is minimally invasive, but carries a risk of malunion and refracture. Rigid antegrade intramedully nailing is possible in adolescents of acceptable size, but has a risk of avascular necrosis. Flexible nailing is minimally invasive and well suited to fractures of the central 2/3 of the diaphysis. In comminuted fractures, it may require supplemental external support. Plate fixation is stable and addresses the entire length of the femur. Soft tissue concerns due to surgical exposure can be minimized by the use of submuscular placement technique.

Multi-germ layer lineage central nervous system repair: nerve and vascular cell generation by embryonic stem cells transplanted in the injured brain.

OBJECT: To restore proper function to a damaged central nervous system (CNS) through transplantation, it is necessary to replace both neural and nonneural elements that arise from different germ layers in the embryo. Mounting evidence indicates the importance of signals related to vasculogenesis in governing neural proliferation and differentiation in early CNS development. Here, the authors examined whether embryonic stem cell (ESC)-derived progenitors can selectively generate both neural and endothelial cells after transplantation in the damaged CNS. METHODS: Injections of 20 nmol N-methyl-D-aspartate created a unilateral striatal injury in 7-day-old rats. One week postinjury, murine ESCs, neural-induced with retinoic acid, were transplanted into the injured striatum. Histological staining, laser confocal microscopy, and transmission electron microscopy of grafted ESCs were performed 1 week posttransplantation. CONCLUSIONS: Transplanted ESCs differentiated into neural cells, which segregated into multiple pools and formed neurons that conformed to host cytoarchitecture. The ESCs also generated endothelial cells, which integrated with host cells to form chimeric vasculature. The combination of ESC pluripotentiality and multiple germ layer differentiation provides a new conceptual framework for CNS repair.

Double lox targeting for neural cell transgenesis.

ES cells differentiated along the neural lineage in vitro are an attractive model system. Here we have developed ES cell lines that are suitable for inserting transgenes at a single chromosomal site. ES cell line CE1 (for Cassette Exchange) contains one "acceptor" module (CE1) that allows for efficient double lox targeting. The site is also permissive for gene expression in neural progenitor cells, as well as differentiated neurons and glia. Line CE2 was derived by swapping a puromycin resistance cassette into CE1. Neural progenitors derived from this line are puromycin-resistant. A beta-actin/GFP expression cassette was inserted into the CE1 site to create CE3. The CE3 cell line was differentiated into neural cells and displayed strong EGFP expression in neural progenitors, differentiated neurons and glia. Differentiated CE3 ES cells (4-/4+ RA) were transplanted into the injured rat somatosensory cortex where many of the transplanted cells survived and differentiated into neurons expressing GFP. This strategy for creating sets of transgenic lines with multiple cassettes inserted into a single chromosomal site provides a powerful tool for studying development and function of ES cell-derived neural cells. Many of these will be useful in transplantation research.