Bio-Templated Growth of Bone Minerals from Modified Simulated Body Fluid on Nanofibrous Decellularized Natural Tissues

Small intestine submucosal (SIS) membrane used in this study is a decellularized, naturally occurring nanofibrous scaffold derived from a submucosal layer of porcine small intestine. It is predominantly composed of type I collagen fibers. Here we studied the bio-templated growth of hydroxylapatite (HAP) bone minerals on the SIS membrane from a modified simulated body fluid (1.5 SBF) at the body temperature, namely, under a near-physiological condition, in order to evaluate its bone bioactivity, the capability of the membrane in bonding with bone tissue once implanted in vivo. Minute HAP crystals were successfully nucleated on the SIS membranes from 1.5 SBF at the body temperature. The crystals were preferentially nucleated along the collagen fibers constituting the SIS membranes. HAP was the major crystalline mineral phase formed during the whole period of time and a minor crystalline phase of tricalcium phosphate (TCP) appeared after the membranes were incubated for 96 h. We also found that the mineralization for 8 h most significantly promoted the osteogenic differentiation of rat mesenchymal stem cells (MSCs) by evaluating the formation of osteogenic markers in MSCs including alkaline phosphatase (early stage marker) as well as osteocalcin and osteopontin (late stage markers). Hence, SIS membranes show excellent bone bioactivity and once mineralized, can significantly promote the osteogenic differentiation of MSCs.