Bone Mineralization Test

Bone Mineralization

As the name suggests, a bone mineralization assay is a method to test the ability of an oil or cream-based product to strengthen the process of bone formation. The construction of a bone starts with the laying down of an extra-cellular matrix facilitated by special fibroblast cells known as osteoblasts. Derived from mesenchymal stem cells (MSCs), osteoblasts secrete & form a mineralized bone matrix containing collagen, calcium, phosphorus, etc. that serve as the base for bone development.
We test all our formulations on MG63 fibroblast cell lines to check their bone formation effect.

What Is MG63 Cell Line & Why We Use It?

MG63 fibroblast cell line has been derived from the bone of a 14-year-old white male suffering from osteosarcoma (bone cancer). These cell lines are an excellent tool for investigating the bone formation effect of our products as they retain and mimic the physiological, biological & developmental properties of osteoblasts in an in vitro (testing in a lab) environment. Moreover, they grow and divide at a much faster rate in comparison to normal osteoblast cell lines.

How We Test The Bone Mineralization Effect Of Our Products?

Step 1

Osteoblast Differentiation & Matrix Mineralization

The first step involves supplementing a set of fibroblast cells (MG63) with osteoblast growth media for 7 days. With exposure to essential osteogenic (bone-formation) factors, these cells initiate the process of bone mineralization.The cells start to grow and secrete collagen, fibronectin & other structural or cell-signaling proteins. Once their growth cycle completes, the concentration of minerals at their surface increases, followed by their deposition onto the matrix. Simultaneously, we introduce the test product & a cell media containing essential growth factors to another set of fibroblast cells for 7 days. With exposure to the test product, the cells may or may not undergo similar events that occur in the first step.

Step 2

Assessment Of Bone Formation By Alizarin-red Staining

We track the level of calcium deposition in both sets of cell lines by introducing the Alizarin-red S stain. Alizarin reacts with the calcium cations and produces a bright red color.The amount of color produced is quantified using a spectrophotometer(an instrument that measures the intensity of light absorbed after it passes through the sample).

What's The Final Result?

For a product to positively affect the bone formation process, itslevel of calcium deposition should be higher than the untreated control. At Areoveda, the tested products reveal similar results!