MSCs have long promised benefits of synthesising bone/cartilage, treating non-unions and potentially accelerating fracture repair. This potential has been tempered by MSC scarcity in the ‘gold-standard’ iliac crest bone marrow aspirate (ICBMA) and the resulting need to expand numbers via cell-culture. Culture of MSCs is time-consuming, expensive and results in cells with a reduced differentiation capacity. The reamer-irrigator-aspirator (RIA) is an innovation designed to reduce intra-medullary (IM) pressures during reaming of long-bones via continuous irrigation and suction. Aspirated contents are passed via a coarse filter, which traps bony-fragments before moving into a ‘waste’ bag - from which MSCs have been previously isolated. We examined liquid and solid phases found in this ‘waste’, performed a novel digestion of the solid phase and made a comparative assessment in terms of number, phenotype and differentiation capacity with matched ICBMA. The filtrate ‘waste’ bag from RIA reaming (6 patients) was filtered (70μm) and the solid fraction digested for 60min (37°C) with collagenase. MSCs were isolated from liquid & solid fractions and from 10ml matched ICBMA. Enumeration of MSCs was achieved via colony-forming-unit-fibroblast (CFUF) assay and flow-cytometry on fresh sample using CD45low, CD271+. MSCs were cultured by virtue of their plastic adherence and passaged in standard, non-haematopoietic media. Passage (P2) cells were differentiated towards osteogenic, adipogenic and chondrogenic lineages with their phenotype assessed with flow cytometry CD33 CD34 CD45 CD73 CD90 CD105.Introduction
Methods
Current literature supports the use of total hip replacement (THR) for the treatment of displaced intra-capsular proximal femoral fractures (DIPFF). Case series of patients receiving this treatment show dislocation rates higher than that of patients who have THR to treat osteoarthritis. Large diameter THR have mechanical advantages in terms of dislocation and their role in PFF has yet to be assessed. To assess the role of large-diameter total hip replacements on the rate of dislocation when used to treat displaced intra-capsular proximal femoral fractures. Design: Single surgeon, case series Setting: Level I trauma centre Inclusion criteria: Displaced intra-capsular proximal femoral fracture (Garden III & IV). Independently mobile pre-operatively for distances greater than a mile, with no more than 1 stick as a mobility aid. Abbreviated mental test score of 9/10 or greater Exclusion criteria: Patient under 60 Pathological fractures Additional fractures of the femur Outcomes Mortality Morbidity (Including dislocation) Oxford Hip Score SF12 Patients/Participants: Retrospective study to assess patients who presented between May 2006 and December 2008 and met the requirements had a CPTÒ (Zimmer) cemented femoral stem, using 3rd generation cementation techniques, and large diameter Duronò (Zimmer) head and cup (uncemented) inserted as a primary procedure via a modified Hardinge technique. Follow up was via routine clinic appointments, letter to GP and phone conversation with patient.Introduction
Objectives
Iliac crest bone marrow aspirate (ICBMA) is frequently cited as the ‘gold-standard’ source of MSCs. Mesenchymal stem cells have been shown to reside within the intramedullary (IM) cavities of long-bones and a comparative assessment with ICBMA has not yet been performed. Aspiration of the IM cavities of 6 patients' femurs with matched ICBMA was performed. The long-bone-fatty-bone-marrow (LBFBM) aspirated was filtered (70μm) and the solid fraction digested for 60min (37°C) with collagenase. Enumeration was performed via the colony-forming-unit-fibroblast (CFU-F) assay and using the CD45low CD271+ phenotype via flow-cytometry. Passaged (P2) cells were differentiated towards osteogenic, adipogenic and chondrogenic lineages with their phenotype assessed using flow-cytometry CD33 CD34 CD45 CD73 CD90 CD105.Introduction
Methods
Iliac crest bone marrow aspirate (ICBMA) is frequently cited as the ‘gold-standard’ source of MSCs. MSCs have been shown to reside within the intramedullary (IM) cavities of long-bones [Nelea, 2005] however a comparative assessment with ICBMA has not yet been performed and the phenotype of the latter compartment MSCs remains undefined in their native environment. Aspiration of the IM cavities of 6 patients' femurs with matched ICBMA was performed. The long-bone-fatty-bone-marrow (LBFBM) was filtered (70μm) to separate liquid and solid fractions and the solid fraction was briefly (60min, 37oC) digested with collagenase. MSC enumeration was performed using the colony-forming-unit-fibroblast (CFU-F) assay and quantification of cells with the CD45low CD271+ phenotype by flow-cytometry. [Jones 2002, Buhring 2007] MSCs were cultured and standard expansion media and passage 2 cells were differentiated towards osteogenic, adipogenic and chondrogenic lineages.Introduction
Methods
Therapeutic exploitation of MSCs in orthopaedics has been tempered by their scarcity within ‘gold-standard’ iliac crest bone marrow aspirate (ICBMA) and the resulting need to expand cells in vitro. This is time-consuming, expensive and results in cells with a reduced differentiation capacity. [Banfi 2000] The RIA is a device that provides continuous irrigation and suction during reaming of long bones. Aspirated contents pass via a filter, trapping bony-fragments, before moving into a ‘waste’ bag, from which MSCs have been previously isolated. [Porter 2009] We hypothesised that ‘waste’ RIA bag contains more MSCs than a standard aspirated volume of ICBMA (30 ml). We further hypothesised than a fatty solid phase within this ‘waste bag’ contains many MSCs trapped within the adipocyte-rich stromal network and hence requiring an enzymatic digestion for their efficient release [Jones 2006]. The discarded filtrate ‘waste’ bag that contained saline from marrow cavity irrigation procedure from RIA reaming (7 patients) was filtered (70μm) and the solid fraction digested for 60min (37oC) with collagenase. MSC enumeration was performed using the colony-forming-unit-fibroblast (CFU-F). Following culture in standard expansion media, passage 2 cells were differentiated towards osteogenic, adipogenic and chondrogenic lineages and their phenotype was assessed using flow cytometry. ICBMA from the same patients was used as controls.Introduction
Methods
Femoral neck stress fractures (FNSF) are uncommon, representing around 5% of all stress fractures. In military personnel, FNSF represents one of the severest complications of military training, which can result in medical discharge. Clinical examination findings are frequently non-specific and plain radiography may be inconclusive leading to missed or late diagnosis of FNSF. This paper highlights the significance of FNSF’s in military personnel and alerts physicians to the potential diagnosis. We identified all military recruits, aged 17 to 26, who attended the Infantry Training Centre (Catterick, UK), over a four-year period from the 1st July 2002 to 30th June 2006, who suffered a FNSF. The medical records, plain radiographs, bone scans and MRI’s of the recruits were retrospectively reviewed. Of 250 stress fractures, 20 were of the femoral neck; representing 8% of all stress fractures and an overall FNSF rate of 12 in 10,000 military recruits. FNSF’s were most prevalent amongst Parachute Regiment recruits (1 in 250, p<
0.05). Onset of symptoms was most commonly between 13–16 weeks from the start of training. The majority (17/20, 85%) of FNSF’s were undisplaced, these were all treated conservatively. Three FNSF’s were displaced on presentation and were treated surgically. Overall, the medical discharge rate was 40% (8/20). FNSF’s are uncommon and the diagnosis remains a challenge to clinicians and requires a high index of suspicion in young athletic individuals. In such individuals early referral for MRI is recommended, to aid prompt diagnosis and treatment, to prevent serious sequelae. Correspondence should be addressed to Major M Butler RAMC, Princess Elizabeth Orthopaedic Centre, Royal Devon and Exeter Hospital, Exeter, Devon.
