Platelet lysate can support the development of a 3D-engineered skin for clinical application

Abstract

Safety concerns associated with foetal bovine serum (FBS) have restricted its translation into clinics. We hypothesised that platelet lysate (PL) can be utilised as a safe alternative to produce serum-free 3D-engineered skin. PL supported a short-term expansion of fibroblasts, with negligible replication-induced senescence and directed epidermal stratification. PL-expanded fibroblasts were phenotypically separated into three subpopulations of CD90+FAP+, CD90+FAP− and CD90−FAP+, based on CD90 (reticular marker) and FAP (papillary marker) expression profile. PL drove the expansion of the intermediate CD90+ FAP+ subpopulation in expense of reticular CD90+FAP−, which may be less fibrotic once grafted. The 3D-engineered skin cultured in PL was analysed by immunofluorescence using specific markers. Detection of ColIV and LMN-511 confirmed basement membrane. K10 confirmed near native differentiation pattern of neo-epidermis. CD29- and K5-positive interfollicular stem cells were also sustained. Transmission and scanning electron microscopies detailed the ultrastructure of the neo-dermis and neo-epidermis. To elucidate the underlying mechanism of the effect of PL on skin maturation, growth factor contents in PL were measured, and TGF-β1 was identified as one of the most abundant. TGF-β1 neutralising antibody reduced the number of Ki67-positive proliferative cells, suggesting TGF-β1 plays a role in skin maturation. Moreover, the 3D-engineered skin was exposed to lucifer yellow on days 1, 3 and 5. Penetration of lucifer yellow into the skin was used as a semi-quantitative measure of improved barrier function over time. Our findings support the concept of PL as a safe and effective serum alternative for bioengineering skin for cell therapies.

A new subtype of artificial cell‐derived vesicles from dental pulp stem cells with the bioequivalence and higher acquisition efficiency compared to extracellular vesicles

“…Compared with plain DMEM medium, ACDVs supplemented DMEM attracted twice more MDFs over 24 h culture (Figure 4b,c). Similar effect was reported previously (Banakh et al., 2023; Sovkova et al., 2018). According to the scratch assay (Figure 4d), comparing with the control, ACDVs formulation showed prominent effect on the migration of fibroblasts early on 12 h after the scratch was done.…”

“…Besides above varied features, we found that ACDVs and EVs were consistent in multiple dimensions of EVs, which echoed previous studies (Bakhtyar et al., 2018; Banakh et al., 2023; Li et al., 2016). TEM, Western Blotting and Nanoparticle tracking analysis were used as the general criteria for EV identification.…”

Recent Achievements in the Development of Biomaterials Improved with Platelet Concentrates for Soft and Hard Tissue Engineering Applications

“…The PL can be utilized as a safe alternative to produce 3D-engineered skin products. PL supports the expansion of fibroblasts, with negligible replication-induced senescence and directed epidermal stratification [43].…”