Abstract
Aims: To substantiate the osteogenic differentiation of human Umbilical cord Mesenchymal Stem Cells (hUMSCs) under the influence of Bone Dust (BD) by detecting expression of genes RUNX2 and IBSP in the differentiated cells.
 Study Design: In vitro study.
 Place and Duration of Study: MDRL Cell Culture Lab Ziauddin University and Dr.Zafar H.Zaidi Institute of Proteomics, University of Karachi, between July 2020 and July 2021.
 Methodology: We designed an innovative experiment to differentiate hUMSCs into osteoblasts under the influence of BD, without using any conventional differentiating medium. The hUMSCs were isolated from umbilical cord and cultured in a 6-well plate with 3μm inserts, in which BD was lodged, preventing direct contact of BD with hUMSCs. The plates with hUMSCs and BD-filled inserts were incubated at 37°C for 14 days after which RNA extraction, cDNA synthesis and qPCR for two genes was performed. Expression of Runt-related transcription factor-2(RUNX2) and Integrin-binding Sialoprotein (IBSP) were quantified.
 Results: Results revealed an increased expression of the late osteogenic marker IBSP(p=0.01) signifying that the hUMSCs had undergone differentiation into mature osteoblasts.
 Conclusion: This novel experiment identifies BD as a new source for osteogenic differentiation of hUMSCs in in vitro experiments.
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