The absorption and transport scattering coefficients of caucasian and negroid dermis, subdermal fat and muscle have been measured for all wavelengths between 620 and 1000 nm. Samples of tissue 2 mm thick were measured ex vivo to determine their reflectance and transmittance. A Monte Carlo model of the measurement system and light transport in tissue was then used to recover the optical coefficients. The sample reflectance and transmittance were measured using a single integrating sphere 'comparison' method. This has the advantage over conventional double-sphere techniques in that no corrections are required for sphere properties, and so measurements sufficiently accurate to recover the absorption coefficient reliably could be made. The optical properties of caucasian dermis were found to be approximately twice those of the underlying fat layer. At 633 nm, the mean optical properties over 12 samples were 0.033 mm(-1) and 0.013 mm(-1) for absorption coefficient and 2.73 mm(-1) and 1.26 mm(-1) for transport scattering coefficient for caucasian dermis and the underlying fat layer respectively. The transport scattering coefficient for all biological samples showed a monotonic decrease with increasing wavelength. The method was calibrated using solid tissue phantoms and by comparison with a temporally resolved technique.

Porous collagen-glycosaminoglycan (collagen-GAG) scaffolds have shown promising clinical results for wound healing; however, these scaffolds do not replace the dermal and epidermal layer simultaneously and rely on local endogenous signaling to direct healing. Functionalizing collagen-GAG scaffolds with signaling factors, and/or additional matrix molecules, could help overcome these challenges. An ideal candidate for this is platelet-rich plasma (PRP) as it is a natural reservoir of growth factors, can be activated to form a fibrin gel, and is available intraoperatively. We tested the factors released from PRP (PRPr) and found that at specific concentrations, PRPr enhanced cell proliferation and migration and induced angiogenesis to a greater extent than fetal bovine serum (FBS) controls. This motivated us to develop a strategy to successfully incorporate PRP homogeneously within the pores of the collagen-GAG scaffolds. The composite scaffold released key growth factors for wound healing (FGF, TGFβ) and vascularization (VEGF, PDGF) for up to 14 days. In addition, the composite scaffold had enhanced mechanical properties (when compared to PRP gel alone), while providing a continuous upper surface of extracellular matrix (ECM) for keratinocyte seeding. The levels of the factors released from the composite scaffold were sufficient to sustain proliferation of key cells involved in wound healing, including human endothelial cells, mesenchymal stromal cells, fibroblasts, and keratinocytes; even in the absence of FBS supplementation. In functional in vitro and in vivo vascularization assays, our composite scaffold demonstrated increased angiogenic and vascularization potential, which is known to lead to enhanced wound healing. Upon pro-inflammatory induction, macrophages released lower levels of the pro-inflammatory marker MIP-1α when treated with PRPr; and released higher levels of the anti-inflammatory marker IL1-ra upon both pro-and anti-inflammatory induction when treated with the composite scaffold. Finally, our composite scaffold supported a co-culture system of human fibroblasts and do Amaral et al. PRP Incorporation Into Collagen-Based Scaffold keratinocytes that resulted in an epidermal-like layer, with keratinocytes constrained to the surface of the scaffold; by contrast, keratinocytes were observed infiltrating the PRPfree scaffold. This novel composite scaffold has the potential for rapid translation to the clinic by isolating PRP from a patient intraoperatively and combining it with regulatory approved scaffolds to enhance wound repair.

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Successful therapeutic strategies for wound healing rely on proper vascularization while inhibiting fibrosis. However, scaffolds designed for skin tissue engineering generally lack the biochemical cues that can enhance their vascularization without inducing fibrosis. Therefore, the objective of this work is to incorporate platelet‐rich plasma (PRP), a natural source of angiogenic growth factors, into a gelatin methacrylate (GelMA) hydrogel, yielding a bioink that can subsequently be used to 3D print a novel regenerative scaffold with defined architecture for skin wound healing. A PRP‐activated bioink is successfully 3D printed, and the resulting scaffolds present similar structural, rheological, and mechanical properties compared to GelMA‐only scaffolds. Furthermore, 3D printed PRP‐activated scaffolds facilitate controlled release of PRP‐derived growth factors for up to 14 days, presenting superior angiogenic potential in vitro (e.g., tubulogenesis assay) and in vivo (chick chorioallantoic membrane) compared to GelMA‐only scaffolds, while not inducing a myofibroblastic phenotype in fibroblasts (e.g., α‐smooth muscle actin expression). This disruptive technology offers the opportunity for a patient's autologous growth factors to be incorporated into a tailored 3D‐printed scaffold in theatre prior to implantation, as part of a single‐stage procedure, and has potential in other tissue engineering applications in which enhanced vascularization with limited fibrosis is desired.

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