Identification and quantification of growth factors involved in growth and development of cartilage present in exosomes of lactating mother’s milk of cleft infant as compared to non-cleft infant – a protocol for an analytical observational study

Abstract

Background: Cleft lip and palate (CLP) have complex etiology and even after successful cleft surgeries in a CLP child, there are chances of relapse as there may be a lack of something at the cellular level. Several growth factors like TGF-β, IGF, and BMP7 are involved in modulating chondrocyte homeostasis and can also induce chondrogenic differentiation in the bone marrow. Objectives: To identify growth factors involved in the growth and development of cartilage present in exosomes of lactating maternal milk of cleft and non-cleft children. Methodology: There will be two groups: GROUP-A: The lactating mother of a CLP child. GROUP-B: The lactating mother of a non-cleft child. Mothers with a CLP child and a non-cleft child between the age of 0–6 months will be selected for the study. The need for the study will be explained to the mothers and consent will be taken. A milk sample of 2ml will be collected in a falcon tube and transported in cold storage facilities to the laboratory. Initially, the isolation of exosomes from milk samples will be done, from which total proteins will be isolated by centrifugation process. This will be followed by an analysis of growth factors from the isolated total proteins (from both exosomes) through immunoblotting. Once the analysis is done, the quality and quantity of growth factors responsible for cartilage growth and development will be compared. Expected results: Isolated maternal exosomes are expected to contain an ample amount of growth factors involved in cartilage growth and development, highlighting its potential for use as a therapy alongside prevalent procedures in CLP. Conclusion: In this study, we expect that maternal exosomes act as a carrier of factors that can have therapeutic significance to the natural compensation of the cartilage to maintain nasal symmetry by naso-alveolar molding.