Fabrication of Helix aspersa Extract Loaded Gradient Scaffold with an Integrated Architecture for Osteochondral Tissue Regeneration: Morphology, Structure, and In Vitro Bioactivity.
Sedef TamburaciMerve PerpelekSelma AydemirBasak BaykaraHasan HavitciogluFunda TihminliogluPublished in: ACS applied bio materials (2023)
Regeneration of osteochondral tissue with its layered complex structure and limited self-repair capacity has come into prominence as an application area for biomaterial design. Thus, literature studies have aimed to design multilayered scaffolds using natural polymers to mimic its unique structure. In this study, fabricated scaffolds are composed of transition layers both chemically and morphologically to mimic the gradient structure of osteochondral tissue. The aim of this study is to produce gradient chitosan (CHI) scaffolds with bioactive snail ( Helix aspersa ) mucus (M) and slime (S) extract and investigate the structures regarding their physicochemical, mechanical, and morphological characteristics as well as in vitro cytocompatibility and bioactivity. Gradient scaffolds (CHI-M and CHI-S) were fabricated via a layer-by-layer freezing and lyophilization technique. Highly porous and continuous 3D structures were obtained and observed with SEM analysis. In addition, scaffolds were physically characterized with water uptake test, micro-CT, mechanical analysis (compression tests), and XRD analysis. In vitro bioactivity of scaffolds was investigated by co-culturing Saos-2 and SW1353 cells on each compartment of gradient scaffolds. Osteogenic activity of Saos-2 cells on extract loaded gradient scaffolds was investigated in terms of ALP secretion, osteocalcin (OC) production, and biomineralization. Chondrogenic bioactivity of SW1353 cells was investigated regarding COMP and GAG production and observed with Alcian Blue staining. Both mucus and slime incorporation in the chitosan matrix increased the osteogenic differentiation of Saos-2 and SW1353 cells in comparison to the pristine matrix. In addition, histological and immunohistological staining was performed to investigate ECM formation on gradient scaffolds. Both characterization and in vitro bioactivity results indicated that CHI-M and CHI-S scaffolds show potential for osteochondral tissue regeneration, mimicking the structure as well as enhancing physical characteristics and bioactivity.
Keyphrases
- tissue engineering
- induced apoptosis
- cell cycle arrest
- stem cells
- drug delivery
- wound healing
- oxidative stress
- bone marrow
- mental health
- systematic review
- high resolution
- mesenchymal stem cells
- magnetic resonance imaging
- computed tomography
- endoplasmic reticulum stress
- platelet rich plasma
- physical activity
- contrast enhanced
- signaling pathway
- cell death
- risk assessment
- transcription factor
- pi k akt
- cell proliferation
- human health
- mass spectrometry
- climate change
- extracellular matrix
- hyaluronic acid
- data analysis
- dna binding