Inductive and conductive effects on bone cells using an in-vitro mouse calvarial model
Embargo Date
2026-05-28
OA Version
Citation
Abstract
INTRODUCTION: This thesis explores findings from three trials utilizing a mouse in-vitro calvarial model. The primary aim was to establish a reproducible in-vitro bone tissue model for testing the inductive and conductive potential of materials for bone regeneration.
MATERIALS AND METHODS: Trial I: Assessed the behavior of normal calvarial bone under laboratory-standard conditions protocol with (DMEM). Histological examinations and micro-CT were analyzed on 10 specimens over 18 days.
Trial II: Evaluated the effects of ascorbic acid (AA) in three different culture media formulae on a critical size defect repair. Histological examinations and alkaline phosphatase (ALP) activity were analyzed on 40 specimens over 21 days.
Trial III: Explored the potential of "DAG" (dexamethasone, ascorbic acid, β-glycerophosphate) as an osteoinductive stimulus using (DMEM). Histological examinations and ALP activity were analyzed on 20 specimens over 36 days. Micro-CT scans were conducted on four samples, one selected from each group.
RESULTS: Trial I: Histological observations demonstrated a significant reduction in cellular activity and thinning of the periosteal layer from day 0 to day 18. Micro-CT analysis indicated an increased bone mineral density over time (n=1 for each time period).
Trial II: Treatment with α-MEM supplemented with 200 mg/L AA led to complete closure of a 2 mm defect, while treatment with α-MEM containing 50 mg/L AA resulted in a dramatic reduction in calvarial bone thickness and cellular density. DMEM with 150 mg/L AA preserved the defect size and bone structure with minimal changes.
Trial III: Histological examinations, ALP activity, and micro-CT analysis revealed elevated levels of cell activity in DAG-treated groups relative to saline controls, with the highest cell proliferation and migration observed in Group 4, treated with OSSIX® Plus and DAG.
CONCLUSIONS: Trial I: Our findings underscore the importance of timely intervention when using this model to test materials with osteogenic potential, to capitalize on the thick periosteal layers present at the time of harvest.
Trial II: AA concentration in the culture medium profoundly impacted bone development. DMEM with 150 AA and a 2 mm defect can be defined as a reproducible critical size defect in an in-vitro mouse calvarial model.
Trial III: DAG induced cell migration and proliferation and has potential for directed induction of bone cell migration.
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Attribution-NonCommercial-NoDerivatives 4.0 International