Bioactive Agents for Functionalization of Biomaterials for Precise Tissue Engineering

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The lack of bioactivity in three-dimensional (3D)-printing of poly-є-caprolactone (PCL) scaffolds limits cell-material interactions in bone tissue engineering. This constraint can be overcome by surface-functionalization using glycosaminoglycan-like anionic polysaccharides, e.g., carboxymethyl cellulose (CMC), a plant-based carboxymethylated, unsulfated polysaccharide, and κ-carrageenan, a seaweed-derived sulfated, non-carboxymethylated polysaccharide. The sulfation of CMC and carboxymethylation of κ-carrageenan critically improve their bioactivity. However, whether sulfated carboxymethyl cellulose (SCMC) and carboxymethyl κ-carrageenan (CM-κ-Car) affect the osteogenic differentiation potential of pre-osteoblasts on 3D-scaffolds is still unknown. Here, we aimed to assess the effects of surface-functionalization by SCMC or CM-κ-Car on the physicochemical and mechanical properties of 3D-printed PCL scaffolds, as well as the osteogenic response of pre-osteoblasts. MC3T3-E1 pre-osteoblasts were seeded on 3D-printed PCL scaffolds that were functionalized by CM-κ-Car (PCL/CM-κ-Car) or SCMC (PCL/SCMC), cultured up to 28 days. The scaffolds’ physicochemical and mechanical properties and pre-osteoblast function were assessed experimentally and by finite element (FE) modeling. We found that the surface-functionalization by SCMC and CM-κ-Car did not change the scaffold geometry and structure but decreased the elastic modulus. Furthermore, the scaffold surface roughness and hardness increased and the scaffold became more hydrophilic. The FE modeling results implied resilience up to 2% compression strain, which was below the yield stress for all scaffolds. Surface-functionalization by SCMC decreased Runx2 and Dmp1 expression, while surface-functionalization by CM-κ-Car increased Cox2 expression at day 1. Surface-functionalization by SCMC most strongly enhanced pre-osteoblast proliferation and collagen production, while CM-κ-Car most significantly increased alkaline phosphatase activity and mineralization after 28 days. In conclusion, surface-functionalization by SCMC or CM-κ-Car of 3D-printed PCL-scaffolds enhanced pre-osteoblast proliferation and osteogenic activity, likely due to increased surface roughness and hydrophilicity. Surface-functionalization by SCMC most strongly enhanced cell proliferation, while CM-κ-Car most significantly promoted osteogenic activity, suggesting that surface-functionalization by CM-κ-Car may be more promising, especially in the short-term, for in vivo bone formation.

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The physico-chemical properties of CaP-based biomaterials affect the activity and function of immune cells, osteoclasts, and osteoblasts. Abbreviation: BCP, biphasic calcium phosphate; TCP, tricalcium phosphate; HA, hydroxyapatite; MSC, mesenchymal stem cell; OCP, octacalcium phosphate.
Mini Review
16 May 2022

Calcium phosphate (CaP)-based bioceramics are the most widely used synthetic biomaterials for reconstructing damaged bone. Accompanied by bone healing process, implanted materials are gradually degraded while bone ultimately returns to its original geometry and function. In this progress report, we reviewed the complex and tight relationship between the bone healing response and CaP-based biomaterials, with the emphasis on the in vivo degradation mechanisms of such material and their osteoinductive properties mediated by immune responses, osteoclastogenesis and osteoblasts. A deep understanding of the interaction between biological healing process and biomaterials will optimize the design of CaP-based biomaterials, and further translate into effective strategies for biomaterials customization.

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(A) In vitro effects of naringin, reproduced with permission (Yu et al., 2020). (B) Schematic diagram of some osteogenic pathways and genes influenced by berberine, and schematic diagram of berberine inhibiting osteoclasts by affecting the binding of RANK and RANKL, reproduced with permission (Zhang et al., 2021a). (C) Schematic diagram of the bone remodeling mechanism and the role of ginsenosides, reproduced with permission (Yang N. et al., 2020). (D) Biological mechanism leading to inhibition of osteoclast differentiation of BMMs by CURCGNPs through RANKL-induced signaling pathways, reproduced with permission (Heo et al., 2014).
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(A) Immunofluorescence staining of TNF-α, TGF-β, and IL-10 in the wounds at Day 3 and Day 7. (B) Quantitative analysis of Immunofluorescence staining of TNF-α, TGF-β, and IL-10 in the wounds at Day 3 and Day 7.
Original Research
09 February 2022

Bacterial infection, inflammatory disorder, and poor angiogenesis of tissue in chronic wounds are the main reasons why wounds are difficult to heal. In this study, a novel MSN-PEG@AS/BP nano-spray was designed to solve these issues. Astragaloside IV (AS) was loaded in mesoporous silica nanoparticles (MSN) to enhance angiogenesis and regulate inflammation, and the two-dimensional (2D) nanosheet black phosphorus (BP) was used to kill bacteria through a photothermal effect. Under thermal decomposition, the covalent bond of polyethylene glycol (PEG) was broken, releasing AS to promote the proliferation of fibroblasts, the formation of blood vessels, and the resolution of inflammation. AS can promote the polarization of the anti-inflammatory (M2) macrophage phenotype to enhance the deposition of extracellular matrix and the formation of blood vessels. Besides, BP showed a significant photothermal effect and nearly 99.58% of Escherichia coli and 99.13% of Staphylococcus aureus were killed in an antibacterial study. This nano-spray would be a novel therapeutic agent for infected wound treatment.

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Frontiers in Bioengineering and Biotechnology

Comprehensive Exploration of Biomaterials and Nanobiotechnology for Tissue Regeneration and Organ Reconstruction
Edited by Weiyan Sun, Yi Hong, Dongxu Ke
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25 June 2025
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