Intrafibrillar mineralization plays a critical role in attaining desired mechanical properties of bone. It is well known that amorphous calcium phosphate (ACP) infiltrates into the collagen through the gap regions, but its underlying driving force is not understood. Based on the authors’ previous observations that a collagen fibril has higher piezoelectricity at gap regions, it was hypothesized that the piezoelectric heterogeneity of collagen helps ACP infiltration through the gap. To further examine this hypothesis, the collagen piezoelectricity of osteogenesis imperfecta (OI), known as brittle bone disease, is characterized by employing Piezoresponse Force Microscopy (PFM). The OI collagen reveals similar piezoelectricity between gap and overlap regions, implying that losing piezoelectric heterogeneity in OI collagen results in abnormal intrafibrillar mineralization and, accordingly, losing the benefit of mechanical heterogeneity from the fibrillar level. This finding suggests a perspective to explain the ACP infiltration, highlighting the physiological role of collagen piezoelectricity in intrafibrillar mineralization.