An extracellular matrix hydrogel from porcine urinary bladder for tissue engineering: In vitro and in vivo analyses

Abstract

BACKGROUND: The necessity to manufacture scaffolds with superior capabilities of biocompatibility and biodegradability has led to the production of extracellular matrix (ECM) scaffolds. Among their advantages, they allow better cell colonization, which enables its successful integration into the hosted tissue, surrounding the area to be repaired and their formulations facilitate placing it into irregular shapes. The ECM from porcine urinary bladder (pUBM) comprises proteins, proteoglycans and glycosaminoglycans which provide support and enable signals to the cells. These properties make it an excellent option to produce hydrogels that can be used in regenerative medicine. OBJECTIVE: The goal of this study was to assess the biocompatibility of an ECM hydrogel derived from the porcine urinary bladder (pUBMh) in vitro using fibroblasts, macrophages, and adipose-derived mesenchymal stem cells (AD-MCSs), as well as biocompatibility in vivo using Wistar rats. METHODS: Effects upon cells proliferation/viability was measured using MTT assay, cytotoxic effects were analyzed by quantifying lactate dehydrogenase release and the Live/Dead Cell Imaging assay. Macrophage activation was assessed by quantification of IL-6, IL-10, IL-12p70, MCP-1, and TNF-α using a microsphere-based cytometric bead array. For in vivo analysis, Wistar rats were inoculated into the dorsal sub-dermis with pUBMh. The specimens were sacrificed at 24 h after inoculation for histological study. RESULTS: The pUBMh obtained showed good consistency and absence of cell debris. The biocompatibility tests in vitro revealed that the pUBMh promoted cell proliferation and it is not cytotoxic on the three tested cell lines and induces the production of pro-inflammatory cytokines on macrophages, mainly TNF-α and MCP-1. In vivo, pUBMh exhibited fibroblast-like cell recruitment, without tissue damage or inflammation. CONCLUSION: The results show that pUBMh allows cell proliferation without cytotoxic effects and can be considered an excellent biomaterial for tissue engineering.

Advancements and Challenges in Hydrogel Engineering for Regenerative Medicine

“…Extracellular matrix (ECM)-derived and decellularized hydrogels, utilizing materials from sources like the porcine urinary bladder (pUBM) [9,49] and decellularized tissues such as the small intestine submucosa (SISMA) [22], offer a biomimetic environment conducive to cell attachment, proliferation, and differentiation.…”

“…The use of extracellular matrix hydrogels from the porcine urinary bladder has highlighted their biocompatibility and biodegradability, which significantly enhance cell colonization and their integration into host tissues. This strategy is designed to effectively tackle the challenges associated with repairing irregularly shaped defects [9]. Additionally, the development of a tissue-adhesive hydrogel through recombinant tyrosinase-mediated crosslinking has been optimized for rapid gelation, enhanced physical properties, and strong adhesive qualities, enabling its use as an injectable and sprayable solution across various applications in regenerative medicine [119].…”

“…The adaptability of hydrogels is further exemplified by their customizability and tunability, enabling the modeling of patient-specific tissues and controlled drug delivery via adjustable erosion rates, showcasing their ability to cater to a wide array of medical requirements [23,37]. Their non-toxic nature, rapid gelation abilities, robust adhesive strength, and minimal immune responses further solidify their suitability and efficacy for various medical applications [9,119].…”

“…These scaffolds provide an optimal environment that supports tissue integration and stem cell development while minimizing potential adverse reactions [1][2][3][4][5][6][7]. Achieving a harmonious balance between the biocompatibility, biodegradability, and mechanical strength of these scaffolds is a significant challenge but is pivotal for their successful clinical application [8][9][10][11]. The development of materials that are both bioactive and biodegradable poses particular difficulties in the regeneration of complex tissues, such as those requiring vascularization [8].…”