Abstract: The present disclosure provides, in various aspects, a method of forming a prefabricated innervated pre-vascularized pre-laminated (PIPP) flap having a stoma or lumen. The method includes providing a cell construct including skin cells and/or mucosa cells. The method further includes forming an integrated in vivo composite at a donor site by grafting the cell construct onto a muscle. The method further includes stabilizing the composite on an obturator component. The method further includes developing a microvascular system in the composite by retaining it in vivo at the donor site for a predetermined period of time. The method further includes removing the obturator component from the stoma or lumen. In certain aspects, the present disclosure also provides a method of restoring a defect including damaged or surgically removed soft tissue using a PIPP flap. In certain aspect, the present disclosure also provides an obturator component for maintaining the stoma or lumen.

Abstract: This disclosure provides a method of determining the viability of at least one cell via quantification of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), flavin adenine dinucleotide (FAD), and/or collagen. The method includes the steps of contacting the at least one cell with light having a wavelength of from 700 to 900 nm, using two photon excitation, or from 335 to 400 nm, using one photon excitation, to induce an optical response from the NAD(P)H, FAD, and/or collagen and measuring the optical response. The method also includes the step of quantifying one or more of an amount, spatial localization, and/or time-dependent response of the NAD(P)H, FAD, and/or collagen utilizing the optical response.