Abstract
BackgroundThe translation from animal research into the clinical environment remains problematic, as animal systems do not adequately replicate the human in vivo environment. Bioreactors have emerged as a good alternative that can reproduce part of the human in vivo processes at an in vitro level. However, in vitro bone formation platforms primarily utilize stem cells only, with tissue based in vitro systems remaining poorly investigated. As such, the present pilot study explored the tissue behavior and cell survival capability within a new in vitro skeletal muscle tissue-based biomaterial organoid bioreactor system to maximize future bone tissue engineering prospects.ResultsThree dimensional printed β-tricalcium phosphate/hydroxyapatite devices were either wrapped in a sheet of rat muscle tissue or first implanted in a heterotopic muscle pouch that was then excised and cultured in vitro for up to 30 days. Devices wrapped in muscle tissue showed cell death by day 15. Contrarily, devices in muscle pouches showed angiogenic and limited osteogenic gene expression tendencies with consistent TGF-ß1, COL4A1, VEGF-A, RUNX-2, and BMP-2 up-regulation, respectively. Histologically, muscle tissue degradation and fibrin release was seen being absorbed by devices acting possibly as a support for new tissue formation in the bioceramic scaffold that supports progenitor stem cell osteogenic differentiation.ConclusionsThese results therefore demonstrate that the skeletal muscle pouch-based biomaterial culturing system can support tissue survival over a prolonged culture period and represents a novel organoid tissue model that with further adjustments could generate bone tissue for direct clinical transplantations.
Highlights
The translation from animal research into the clinical environment remains problematic, as animal systems do not adequately replicate the human in vivo environment
These results demonstrate that the skeletal muscle pouch-based biomaterial culturing system can support tissue survival over a prolonged culture period and represents a novel organoid tissue model that with further adjustments could generate bone tissue for direct clinical transplantations
In the tissue wrapping model, no gene expression data could be generated for the 30-day in vitro β-TCP/HA wrapped in skeletal muscle tissue from rats (Fig. 2i, Fig. 5c), as the tissue became necrotic, gradually losing the original tissue structure with fading of nuclei preventing successful extraction of mRNA to be available for quantitative real-time polymerase chain reactions analysis (Fig. 2a-c, Fig. 3a-d)
Summary
The translation from animal research into the clinical environment remains problematic, as animal systems do not adequately replicate the human in vivo environment. Whilst experimental in vitro and in vivo investigations continue to contribute greatly to deciphering specific criteria in biological sciences, the translation from a functional model to the clinical setting takes an exuberant amount of time and consumes vast resources [4]. This is one of the reasons why bone tissue induction models are not yet used and. The primary objective was to see if the models would support tissue survivability and growth into a custom threedimensional (3D) printed bone inductive biomaterial [34, 35], whereas the secondary objective was to determine if any vasculo−/angiogenic morphogenesis, by monitoring transcriptional and translational markers, would take place as this is a crucial component required for nascent bone tissue formation [20, 21]
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