Acellular Porcine Placental Membranes as a Novel Biomaterial for Tissue Repair Applications

Gustavo Henrique Doná Rodrigues Almeida^1*Luan Stefani Lima¹Mariana Sversut Gibbin²Beatriz Lopomo¹Rafael Oliveira Bergamo¹Raquel Souza da Silva¹Giovanna Vitória Consani Santos¹Bruna Gomes Silva¹Isabela Paulillo D'Onofrio¹Henrique dos Santos²Lediane Silva Pedroso³Tais da Silva³Henrique Lança Fuzeti³Bianca Fuzeti Candian³Thais Naomi Gonçalves Nesiyama⁴João Victor Damin⁵Claudio Guilherme de Assis Oliveira⁵Lucas Paulo Jacinto Saavedra⁵Guilherme Henrique Gonçalves de Almeida⁵Douglas Lopes de Almeida⁵Jaqueline De Carvalho Rinaldi³Francielle Sato²Mauro Luciano Baesso²Luzmarina Hernandes³Flavio Vieira Meirelles⁴Rose Eli Grassi Rici¹Durvanei Augusto Maria¹Paulo Cezar de Freitas Mathias⁵Ana Claudia Oliveira Carreira^1,6

• ¹Postgraduation Program of Anatomy of Domestic and Wild Animals, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, São Paulo, Brazil, São Paulo, Brazil
• ²Department of Physics, State University of Maringá, Maringa, Parana, Brazil
• ³Department of Morphological Sciences, State University of Maringá, Maringá, Paraná, Brazil, Maringá, Brazil
• ⁴Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, Brazil
• ⁵Department of Biotechnology, Genetics and Cell Biology, State University of Maringá, Maringá, Paraná, Brazil, Maringá, Paraná, Brazil
• ⁶Federal University of ABC, Santo André, São Paulo, Brazil

Biological dressings derived from the extracellular matrix (ECM) of human placental tissues have proven effective in treating complex skin wounds and other anatomical sites, offering potential for new therapeutic applications. However, the use of human tissues is limited by ethical and biosafety concerns, restricting large-scale production. To address this, biomaterials from placentas of livestock animals offer a cost-effective, accessible alternative without harming animal welfare. Given pigs' large-scale production, short gestation periods, and abundant material availability, this study aimed to produce, characterize, and validate acellular biomembranes derived from decellularized porcine allantochorion for tissue repair. Placental fragments from Duroc sows were decellularized using a protocol involving immersion and orbital shaking in 0.1% SDS and 0.5% Triton X-100, followed by low-frequency ultrasonication. Accelularity was confirmed by total genomic DNA quantification and H&E and DAPI staining for nuclear visualization. Membrane structure and composition were analyzed using histological, immunohistochemical methods, and scanning electron microscopy. Spectroscopic analyses detected physicochemical changes in placental ECM, and biomechanical testing assessed membrane strength and stiffness. Biological functionality was validated through in vitro cell viability and adhesion assays with canine endothelial progenitor cells and L929 murine fibroblasts. In vivo biocompatibility was tested by subcutaneously implanting the biomaterial in rats for histopathological evaluation. Results showed efficient decellularization, with preserved ECM structure. The scaffolds were cytocompatible, supporting cell adhesion and high viability. In vivo testing revealed no immune rejection, confirming biocompatibility and biodegradability. In conclusion, acellular porcine placental biomembranes have the necessary characteristics to be explored as scaffolds for tissue engineering and novel repair therapies.