Enhanced bone tissue regeneration via mesenchymal stem cells-laden Alginate/GelMa 3D-Bioprinted scaffold treated with Haplophyllum tuberculatum extract

• Published in: Journal of traditional and complementary medicine 2024-11
• Main Authors: Jamshidi-adegani, Fatemeh, Al-Hashmi, Sulaiman, Vakilian, Saeid, Al-kindi, Juhaina, Al-Fahdi, Fahad, Al-Broumi, Mohammed, Al-Hadhrami, Abdullah, Alwahaibi, Nasar, Shalaby, Asem, Gibbons, Simon, Al-Harrasi, Ahmed, Al-Rawahi, Wafa
• Format: Article
• Language: English
• Subjects: Bone; Haplophyllum tuberculatum; Osteogenic differentiation; Stem cells; Tissue engineering
• ISSN: 2225-4110; 2225-4110
• DOI: 10.1016/j.jtcme.2024.11.016

Regeneration of critical-sized bone defects is challenging due to avascular necrosis, inflammation, and disrupted osteogenesis. This study explores the synergistic effects of Haplophyllum tuberculatum (HT) extract and mesenchymal stem cells (MSCs) in an Alg/GelMa 3D-bioprinted scaffold to enhance bone regeneration. Ethanolic extracts of stems (HTS), flowers (HTF), and a mixture (HTM) were obtained via Soxhlet extraction and subsequently characterized. Further investigations into their cytotoxicity, osteogenic and bone tissue regeneration potential were conducted, in vitro and in vivo. Alongside GC-MS analysis, vitamin and elemental analyses of HTS, HTF, and HTM revealed varying concentrations of key elements essential for bone regeneration. All extracts exhibited moderate antioxidant activity, with more potency in the HTF group. Cytotoxicity assays indicated the lack of cytotoxic effects from HTS and HTF on MSCs. The calcium secretion and deposition on MSCs revealed the osteogenic potential of HTF and HTM. Gene and protein expression analyses indicated significant upregulation of osteogenic markers in HTF-treated MSCs. The integration of MSCs into a Alg/GelMa 3D-bioprinted scaffold enhanced their viability and proliferation, validating its biocompatibility. Additionally, HTF promoted calcium deposition and ALP activity in the MSC-laden 3D-bioprinted scaffold. In vivo assessment in a rat calvarial defect model, demonstrated accelerated bone regeneration with HTF-treated scaffolds. The improved bone regeneration, vascularization, and collagen formation in HTF-treated defects were confirmed by histopathology results. This study underscores the significant role of the HTF in promoting osteogenesis, advocating for its incorporation into advanced tissue engineering strategies for bone regeneration. [Display omitted] •HTF extract showed the highest antioxidant activity and osteogenic potential.•Cytotoxicity assays confirmed HTS and HTF extracts are non-toxic to MSCs.•HTF and HTM extracts significantly enhanced calcium secretion and deposition in MSCs.•MSC-laden Alg/GelMa 3D-bioprinted scaffold improved cell viability and proliferation.•HTF-treated scaffolds accelerated bone regeneration and vascularization in vivo.