John Premnath, (2019) Effects of shock waves on osteogenetic potential of mesenchymal stem cells derived from fibrous hamartoma in congenital pseudarthrosis of tibia. Masters thesis, Christian Medical College, Vellore.
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Abstract
OBJECTIVE: To assess the effects of focal shock wave treatment on proliferation and osteogenic differentiation of mesenchymal stem cell derived from fibrous hamartoma in congenital pseudarthrosis of tibia. METHODS: Cryopreserved Mesenchymal stem cells (MSCs) derived from the fibrous hamartoma harvested from 3 patients with congenital pseudarthrosis of tibia(CPT) with clinical features of NF1 were cultured. High energy focal shock waves which were not cytotoxic to CPT MSC was chosen. CPT MSC (n=3) were divided into 3 sets each of control group and shock wave treatment group. The osteoblastic differentiation was monitored for 3 weeks and quantified using gene expression of osteogenic markers (eNOS, ALPL, Osteocalcin, RUNX) using RT PCR and calcium deposition using Alizarin Red S staining. The difference in gene expression was compared using Wilcoxon Signed Ranks Test. RESULTS: CPT MSC had similar phenotypic characteristics of mesenchymal stem cells (CD73+, CD90+, CD105+, CD34-, CD45- and CD14-). No cytotoxicity was observed when CPT MSC were treated with high energy focal shock wave treatment (87%, 89%, 86.4% viable cells). Alizarin red S staining showed a 14% increase in mineralisation of Shock wave treated CPT MSC. The gene expression analysis showed upregulation of eNOS, RUNX2, ALPL and osteocalcin and a down regulation of Osterix-SP7. Our study Suggests there is increase in osteogenic differentiation markers in focal shock wave treated CPT MSCs (not significant). The findings might be clinically significant and requires further investigation. CONCLUSION: The impaired osteogenic potential of fibrous hamartoma cells has been investigated previously. In this study we attempted to use shock wave therapy, a non-invasive modality to improve the osteogenesis of CPT MSC and found upregulation of all associated osteogenic markers at different time points, though this increase in trend did not reach statistical significance because of wide variations known to occur in human primary cells, they might be of clinically relevance. Furthermore, we found that comparing our data with existing studies on pamidronate (in vitro) we found that shock wave treatment may provide a better osteogenic stimulus than the bisphosphonate or rhBMP treatment. Further studies with increased sample size supported by in vivo preclinical animal studies are warranted to establish its clinical significance.
| Item Type: | Thesis (Masters) |
|---|---|
| Uncontrolled Keywords: | Congenital pseudarthrosis of tibia, Focal shock waves, Osteogenetic potential |
| Subjects: | MEDICAL > Orthopaedics |
| Depositing User: | Subramani R |
| Date Deposited: | 01 Sep 2019 09:20 |
| Last Modified: | 01 Sep 2019 09:20 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/11359 |
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