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The Bone & Joint Journal
Vol. 101-B, Issue 3 | Pages 253 - 259
1 Mar 2019
Shafafy R Valsamis EM Luck J Dimock R Rampersad S Kieffer W Morassi GL Elsayed S

Aims

Fracture of the odontoid process (OP) in the elderly is associated with mortality rates similar to those of hip fracture. The aim of this study was to identify variables that predict mortality in patients with a fracture of the OP, and to assess whether established hip fracture scoring systems such as the Nottingham Hip Fracture Score (NHFS) or Sernbo Score might also be used as predictors of mortality in these patients.

Patients and Methods

We conducted a retrospective review of patients aged 65 and over with an acute fracture of the OP from two hospitals. Data collected included demographics, medical history, residence, mobility status, admission blood tests, abbreviated mental test score, presence of other injuries, and head injury. All patients were treated in a semi-rigid cervical orthosis. Univariate and multivariate analysis were undertaken to identify predictors of mortality at 30 days and one year. A total of 82 patients were identified. There were 32 men and 50 women with a mean age of 83.7 years (67 to 100).


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVIII | Pages 60 - 60
1 Sep 2012
Rampersad S Petit A Ruiz JC Wertheimer MR Antoniou J Mwale F
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Purpose

A major drawback of current cartilage and intervertebral disc (IVD) tissue engineering is that human mesenchymal stem cells (MSCs) from osteoarthritic (OA) patients express high levels of type X collagen. Type X collagen is a marker of late stage chondrocyte hypertrophy, linked with endochondral ossification, which precedes bone formation. However, it has been shown that a novel plasma-polymer, called nitrogen-rich plasma-polymerized ethylene (PPE:N), is able to inhibit type X collagen expression in committed MSCs. The aim of this study was to determine if the decreased expression of type X collagen, induced by the PPE:N surfaces is maintained when MSCs are removed from the surface and transferred to pellet cultures in the presence of serum and growth factor free chondrogenic media.

Method

Human MSCs were obtained from aspirates from the intramedullary canal of donors undergoing total hip replacement for OA. Cells were expanded for 2–3 passages and then cultured on polystyrene dishes and on two different PPE:N surfaces: high (H) and low (L) pressure deposition. Cells were transferred for 7 additional days in chondrogenic serum free media (DMEM high glucose supplemented with 2 mM L-glutamine, 20 mM HEPES, 45 mM NaHCO3, 100 U/ml penicillin, 100 ug/ml streptomycin, 1 mg/ml bovine serum albumin, 5 ug/ml insulin, 50 ug/ml ascorbic acid, 5 ng/ml sodium selenite, 5 ug/ml transferrin) in pellet culture or on PS cell culture dishes. RNA was extracted using a standard TRIzol protocol. RT-PCR was realized using Superscript II (RT) and Taq polymerase (PCR) with primers specific for type I and X collagen. GAPDH was used as a housekeeping gene and served to normalize the results.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 563 - 564
1 Nov 2011
Rampersad S Petit A Yao G St-Georges-Robillard A Ruiz J Wertheimer MR Antoniou J Mwale F
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Purpose: Several studies have been directed toward using mesenchymal stem cells (MSCs) from osteoarthritic (OA) patients for cartilage or disc repair because these patients are the ones that will require a source of autologous stem cells if biological repair of tissue lesions is to be a therapeutic option. A major drawback of current cartilage and intervertebral disc tissue engineering repair is that these cells rapidly express type X collagen, a marker of late stage chondrocyte hyperthrophy implicated in endochondral ossification. However, a novel plasma-polymerized thin film material, named nitrogen-rich plasma-polymerized ethylene (PPE:N), is able to inhibit type X collagen expression in committed MSCs. The specific aim of this study was to determine if the suppression of type X collagen by PPE:N is maintained when MSCs are transferred to pellet cultures in chondrogenic defined media.

Method: MSCs were obtained from aspirates from the intramedullary canal of donors undergoing total hip replacement for OA using a protocol approved by the Research Ethics Committee of our institution. Cells were then expanded for 2–3 passages in DMEM high glucose supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin, and finally cultured on polystyrene (PS) cell culture dishes or PPE: N surfaces for 3 and 7 days. Cells were transferred for 3 additional days in a chondrogenic serum free media (DMEM high glucose supplemented with 2 mM L-glutamine, 20 mM HEPES, 45 mM NaHCO3, 100 U/ml penicillin, 100 μg/ml streptomycin, 1 mg/ml bovine serum albumin, 5 μg/ml insulin, 50 μg/ml ascorbic acid, 5 ng/ ml sodium selenite, 5 μg/ml transferrin) in pellet culture or on PS cell culture dishes. Cells were then lysed and proteins were separated on 4–20% acrylamide gels and transferred to nitrocellulose membranes. Type X collagen was detected by Western blot; GAPDH expression was used as an internal control for protein loading.

Results: Results showed that type X collagen protein was expressed in MSCs from OA patients cultured on polystyrene but was suppressed when cultured on PPE: N. Since defined chondrogenic medium are commonly used in pellet culture to promote in vitro chondrogenesis, we then investigated the effect of transferring cells pre-cultured on PPE:N into pellet culture on type X collagen expression. However, the decreased type X collagen expression was not maintained in these conditions and that the expression returned to control values. The decreased type X collagen expression was maintained when the cells were cultured on PS cell culture dishes.

Conclusion: The use of MSCs is promising for tissue engineering of cartilage and intervertebral disc. The present study confirmed the potential of PPE:N surfaces in suppressing type X collagen expression in MSCs from OA patients. However, when MSCs stem cells are transferred to pellet cultures, type X collagen is rapidly re-expressed suggesting that pellet cultures may not be suitable for chondrogenesis of MSCs from OA patients.