Garlic extract enhances bioceramic bone scaffolds through upregulating ALP & BGLAP expression in hMSC-monocyte co-culture.

Bone homeostasis is predicated by osteoblast and osteoclast cell cycles where gene expressions are responsible for their differentiation from human mesenchymal stem cells (hMSC) and monocytes, respectively. The pro-osteogenic potential of an hMSC-monocyte co-culture can be measured through complementary DNA (mRNA synthesis) within the nucleus, known as quantitative polymerase chain reaction (qPCR). Through this technique, the effects of garlic extract (allicin) release from calcium phosphate bone scaffolds on gene expression of bone forming and bone remodeling cells was explored. Results show this complex biomaterial system enhances hMSC differentiation through the upregulation of bone-forming proteins. Osteoblastic gene markers alkaline phosphatase (ALP) and osteocalcin (BGLAP), are respectively upregulated by 3-fold and 1.6-fold by day 14. These mature osteoblasts then upregulate the receptor activator of nuclear factor-kB ligand (RANKL) which recruits osteoclast cells, as captured by a nearly 2-fold higher osteoclast expression of tartrate-resistance acid-phosphatase (ACP5). This also activates antagonist osteoprotegerin (OPG) expression in osteoblasts, decreasing osteoclast resorption potential and ACP5 expression by day 21. The pro-osteogenic environment with garlic extract release is further quantified by a 4× increase in phosphatase activity and visibly captured in immunofluorescent tagged confocal images. Also corroborated by enhanced collagen formation in a preliminary in vivo rat distal femur model, this work collectively reveals how garlic extract can enhance bioceramic scaffolds for bone tissue regenerative applications.

Enhanced osteogenesis of 3D printed ß-TCP scaffolds with Cissus Quadrangularis extract-loaded polydopamine coatings.

Growing demand in bone tissue replacement has shifted treatment strategy from pursuing traditional autogenous and allogeneic grafts to tissue replacement with bioactive biomaterials. Constructs that exhibit the ability to support the bone structure while encouraging tissue regeneration, integration, and replacement represent the future of bone tissue engineering. The present study aimed to understand the osteogenic and mechanical effects of binder jet 3D printed, porous ß-tricalcium phosphate scaffolds modified with a natural polymer/drug coating of polydopamine and Cissus Quadrangularis extract. Compression testing was used to determine the effect the polydopamine coating process had on the mechanical strength of the scaffolds. 3D printed scaffolds with and without polydopamine coatings fractured at 3.88 ± 0.51 MPa and 3.84 ± 1 MPa, respectively, suggesting no detrimental effect on strength due to the coating process. The osteogenic potential of the extract-loaded coating was tested in vitro, under static and dynamic flow conditions, and in vivo in a rat distal femur model. Static osteoblast cultures indicated polydopamine-coated samples with and without the extract exhibited greater proliferation after 3 days (p < 0.05). Similarly, polydopamine resulted in increased proliferation and alkaline phosphatase expression under dynamic flow, but alkaline phosphatase expression was significantly enhanced (p < 0.05) only in samples treated with the extract. Histological analysis of implanted scaffolds showed substantially more new bone growth throughout the implant pores at 4 weeks post-op in polydopamine and extract-loaded implants compared to pure ß-tricalcium phosphate. These results indicated that implants coated with polydopamine and Cissus Quadrangularis extract facilitated osteoblast proliferation and alkaline phosphatase production and improved early bone formation and ingrowth while maintaining mechanical strength.

Effects of MgO and SiO2 on Plasma-Sprayed Hydroxyapatite Coating: An in Vivo Study in Rat Distal Femoral Defects.

Plasma-sprayed hydroxyapatite (HA)-coated titanium implants have been widely used in orthopedic applications due to their inheritance of an excellent mechanical property from titanium and great osteoconductivity from HA. However, the lack of osteoinductivity limits their further applications. In this study, 1 wt % MgO and 0.5 wt % SiO2 were mixed with HA for making plasma-sprayed coatings on titanium implants. Plasma-sprayed HA- and MgO/SiO2-HA-coated titanium implants showed adhesive bond strengths of 25.73 ± 1.92 and 23.44 ± 2.89 MPa, respectively. The presence of MgO and SiO2 significantly increased the osteogenesis, osseointegration, and bone mineralization of HA-coated titanium implants by the evaluation of their histomorphology after 6, 10, and 14 weeks of implantation in rat distal femoral defects. Implant pushout tests also showed a shear modulus of 149.83 ± 3.69 MPa for MgO/SiO2-HA-coated implants after 14 weeks of implantation, compared to 52.68 ± 10.41 MPa for uncoated implants and 83.92 ± 3.68 MPa for pure HA-coated implants; These are differences in the shear modulus of 96% and 56.4%, respectively. This study assesses for the first time the quality of the bone-implant interface of induction plasma-sprayed MgO and SiO2 binary-doped HA coatings on load-bearing implants compared to bare titanium and pure HA coatings in a quantitative manner. Relating the osseointegration and interface shear modulus to the quality of implant fixation is critical to the advancement and implementation of HA-coated orthopedic implants.