Xin Xiong

Bone tissue engineering has emerged as a promising approach to regenerate bone tissue, and injectable biomaterials have shown potential for bone regeneration applications due to their ease of administration and ability to fill irregularly shaped defects. This study aims to develop and characterize an injectable composite material comprising biphasic bone substitutes (BBS) and crosslinked porcine collagen type I for bone regeneration applications. The collagen is crosslinked via a UVA‐riboflavin crosslinking strategy and evaluated by testing the physicochemical properties, including the rheological behavior, dynamic storage modulus ( G′ ) and loss modulus ( G″ ), and in vitro degradation process. The results show that the crosslinked collagen (xCol) exhibits suitable physicochemical properties for injectability and improved viscoelasticity and degradation resistance. Furthermore, xCol is then combined with BBS in a predetermined ratio, obtaining the injectable composite material. The biocompatibility of the materials is evaluated in vitro by XTT and BrdU assays on fibroblasts and preosteoblasts. The results demonstrate that the composite material is biocompatible and supporting pre‐osteoblasts proliferation. In conclusion, the injectable composite material BBS‐xCol has promising physiochemical and biological properties for bone regeneration applications. Further studies are warranted to evaluate its efficacy in vivo and optimize its composition for clinical translation.

Cytocompatibility analyses of new implant materials or biomaterials are not only prescribed by the Medical Device Regulation (MDR), as defined in the DIN ISO Norm 10993-5 and -12, but are also increasingly replacing animal testing. In this context, jellyfish collagen has already been established as an alternative to mammalian collagen in different cell culture conditions, but a lack of knowledge exists about its applicability for cytocompatibility analyses of biomaterials. Thus, the present study was conducted to compare well plates coated with collagen type 0 derived from Rhizostoma pulmo with plates coated with bovine and porcine collagen. The coated well plates were analysed in vitro for their cytocompatibility, according to EN ISO 10993-5/−12, using both L929 fibroblasts and MC3T3 pre-osteoblasts. Thereby, the coated well plates were compared, using established materials as positive controls and a cytotoxic material, RM-A, as a negative control. L929 cells exhibited a significantly higher viability (#### p < 0.0001), proliferation (## p < 0.01), and a lower cytotoxicity (## p < 0.01 and # p < 0.05)) in the Jellagen® group compared to the bovine and porcine collagen groups. MC3T3 cells showed similar viability and acceptable proliferation and cytotoxicity in all collagen groups. The results of the present study revealed that the coating of well plates with collagen Type 0 derived from R. pulmo leads to comparable results to the case of well plates coated with mammalian collagens. Therefore, it is fully suitable for the in vitro analyses of the cytocompatibility of biomaterials or medical devices.