MgCa-Based Alloys Modified with Zn- and Ga-Doped CaP Coatings Lead to Controlled Degradation and Enhanced Bone Formation in a Sheep Cranium Defect Model.

Mg-based biodegradable metallic implants are gaining increased attraction for applications in orthopedics and dentistry. However, their current applications are hampered by their high rate of corrosion, degradation, and rapid release of ions and gas bubbles into the physiological medium. The aim of the present study is to investigate the osteogenic and angiogenic potential of coated Mg-based implants in a sheep cranial defect model. Although their osteogenic potential was studied to some extent, their potential to regenerate vascularized bone formation was not studied in detail. We have studied the potential of magnesium-calcium (MgCa)-based alloys modified with zinc (Zn)- or gallium (Ga)-doped calcium phosphate (CaP) coatings as a strategy to control their degradation rate while enhancing bone regeneration capacity. MgCa and its implants with CaP coatings (MgCa/CaP) as undoped or as doped with Zn or Ga (MgCa/CaP + Zn and MgCa/CaP + Ga, respectively) were implanted in bone defects created in the sheep cranium. MgCa implants degraded faster than the others at 4 weeks postop and the weight loss was ca. 50%, while it was ca. 15% for MgCa/CaP and <10% in the presence of Zn and Ga with CaP coating. Scanning electron microscopy (SEM) analysis of the implant surfaces also revealed that the MgCa implants had the largest degree of structural breakdown of all the groups. Radiological evaluation revealed that surface modification with CaP to the MgCa implants induced better bone regeneration within the defects as well as the enhancement of bone-implant surface integration. Bone volume (%) within the defect was ca. 25% in the case of MgCa/CaP + Ga, while it was around 15% for undoped MgCa group upon micro-CT evaluation. This >1.5-fold increase in bone regeneration for MgCa/CaP + Ga implant was also observed in the histopathological examination of the H&E- and Masson's trichrome-stained sections. Immunohistochemical analysis of the bone regeneration (antiosteopontin) and neovascularization (anti-CD31) at the defect sites revealed >2-fold increase in the expression of the markers in both Ga- and Zn-doped, CaP-coated implants. Zn-doped implants further presented low inflammatory reaction, notable bone regeneration, and neovascularization among all the implant groups. These findings indicated that Ga- and Zn-doped CaP coating is an important strategy to control the degradation rate as well as to achieve enhanced bone regeneration capacity of the implants made of Mg-based alloys.

Zn doped CaP coatings used for controlling the degradation rate of MgCa1 alloy: In vitro anticorrosive properties, sterilization and bacteria/cell-material interactions.

The purpose of this study was to investigate the effect of Zn doped CaP coatings prepared by micro-arc oxidation method, as a possible approach to control MgCa1 alloy degradation. All the prepared coatings comprised a calcium deficient CaP phase. The control in this evaluation was performed with undoped CaP coating in SBF solution at body temperature (37 ± 0.5°C). The investigation involved determination of microchemical, mechanical, morphological, properties along with anticorrosive, cytocompatibility and antibacterial efficacy. The effect of sterilization process on the properties of the surfaces was also investigated. The results showed that the addition of Zn into CaP increased the corrosion resistance of MgCa1 alloy. Moreover, the adhesion strength of the coatings to MgCa1 alloy was enhanced by Zn addition. In cytotoxicity testing of the samples, extracts of the samples in MEM were incubated with L929 cells and malformation, degeneration and lysis of the cells were examined microscopically after 72 h. The results showed that all samples were cytocompatible. The degradation of MgCa1 alloy in the simulated body fluids (SBF) or DMEM was decreased by coating with CaP. Moreover, the degradation rate of CaP was further decreased by adding a small amount of Zn into the CaP matrix. The samples having CaP coatings and Zn doped CaP coating demonstrated antibacterial efficacy against E.coli. As a result, coating of magnesium alloy with Zn-doped CaP decreased the degradation rate, increased the corrosion resistance, cytocompatibility and the antibacterial effects of the alloys.

Potential of pectin for biomedical applications: a comprehensive review.

Pectin is a polysaccharide extracted from various plants, such as apples, oranges, lemons, and it possesses some beneficial effects on human health, including being hypoglycemic and hypocholesterolemic. Therefore, pectin is used in various pharmaceutical and biomedical applications. Meanwhile, its low mechanical strength and fast degradation rate limit its usage as drug delivery devices and tissue engineering scaffolds. To enhance these properties, it can be modified or combined with other organic molecules or polymers and/or inorganic compounds. These materials can be prepared as nano sized drug carriers in the form of spheres, capsules, hydrogels, self assamled micelles, etc., for treatment purposes (mostly cancer). Different composites or blends of pectin can also be produced as membranes, sponges, hydrogels, or 3D printed matrices for tissue regeneration applications. This review is concentrated on the properties of pectin based materials and focus especially on the utilization of these materials as drug carriers and tissue engineering scaffolds, including 3D printed and 3D bioprinted systems covering the studies in the last decade and especially in the last 5 years.

pH responsive release of curcumin from photocrosslinked pectin/gelatin hydrogel wound dressings.

The aim of this study was to develop hydrogel wound dressings made of photocrosslinkable pectin and gelatin with pH dependent release of curcumin, an antimicrobial agent. Methacrylated forms of pectin and gelatin (PeMA and GelMA, respectively) were synthesized, and hydrogels were prepared with different compositions (1:1, 1:2 and 1:3 v/v ratios of PeMA and GelMA) by UV exposure. Pure GelMA was used as control group. Average pore diameter of hydrogels with the highest PeMA content (P1:G1) was 43 µm. All hydrogels showed about 90% swelling. P1:G3 demonstrated the highest stability (retained about 37% of their initial weight after 21 days incubation in PBS), a reasonable compressive modulus (ca. 22 kPa), oxygen permeability (7.44 mg/mL) and preventing ability for bacterial penetration. Therefore, P1:G3 hydrogels were chosen and loaded with curcumin for further studies. In aqueous medium (10 mM PBS, pH 7.4), about 4 times faster release of curcumin was observed than that in medium with pH 5.0. Since infected wounds have alkaline pH compared to healthy tissue, faster release at basic medium is preferable for wound grafts. Disk diffusion tests proved antibacterial efficacy of the hydrogels against S. aureus and E. coli. Live/Dead and Alamar blue assays conducted with L929 fibroblasts showed cytocompatibility of the hydrogels. It was concluded that curcumin loaded P1:G3 hydrogels are promising candidates as wound dressing materials to be further tested in the treatment of infected and chronic wounds.

3D printed hybrid bone constructs of PCL and dental pulp stem cells loaded GelMA.

Fabrication of scaffolds using polymers and then cell seeding is a routine protocol of tissue engineering applications. Synthetic polymers have adequate mechanical properties to substitute for some bone tissue, but they are generally hydrophobic and have no specific cell recognition sites, which leads to poor cell affinity and adhesion. Some natural polymers, have high cell affinity but are mechanically weak and do not have the strength required as a bone supporting material. In the present study, 3D printed hybrid scaffolds were fabricated using PCL and GelMA carrying dental pulp stem cells (DPSCs), which is printed in the gaps between the PCL struts. This cell loaded GelMA was shown to support osteoinductivity, while the PCL provided mechanical strength needed to mimic the bone tissue. 3D printed PCL/GelMA and GelMA scaffolds were highly stable during 21 days of incubation in PBS. The compressive moduli of the hybrid scaffolds were in the range of the compressive moduli of trabecular bone. DPSCs were homogeneously distributed throughout the entire hydrogel component and exhibited high cell viability in both scaffolds during 21 days of incubation. Upon osteogenic differentiation DPSCs expressed two key matrix proteins, osteopontin and osteocalcin. Alizarin red staining showed mineralized nodules, which demonstrates osteogenic differentiation of DPSCs within GelMA. This construct yielded a very high cell viability, osteogenic differentiation and mineralization comparable to cell culture without compromising mechanical strength suitable for bone tissue engineering applications. Thus, 3D printed, cell loaded PCL/GelMA hybrid scaffolds have a great potential for use in bone tissue engineering applications.

3D printed poly(ε-caprolactone) scaffolds modified with hydroxyapatite and poly(propylene fumarate) and their effects on the healing of rabbit femur defects.

A large variety of approaches have been used to treat large and irregular shaped bone defects with less than optimal success due to material or design issues. In recent years patient specific constructs prepared by additive manufacturing provided a solution to the need for shaping implants to fit irregular defects in the surgery theater. In this study, cylindrical disks of poly(ε-caprolactone) (PCL) were printed by fused deposition modeling and modified with nanohydroxyapatite (HAp) and poly(propylene fumarate) (PPF) to create a mechanically strong implant with well-defined pore size and porosity, controllable surface hydrophilicity (with PPF) and osteoconductivity (with HAp). Cytotoxicity, irritation and inflammation tests demonstrated that the scaffolds were biocompatible. PCL/HAp and PCL/HAp/PPF scaffolds were implanted in the femurs of rabbits with and without seeding with rabbit Bone Marrow Stem Cells (BMSC) and examined after 4 and 8 weeks with micro-CT, mechanically and histologically. BMSC seeded PCL/HAp/PPF scaffolds showed improved tissue regeneration as determined by bone mineral density and micro-CT. Compressive and tension stiffness values (394 and 463 N mm-1) were significantly higher than those of the healthy rabbit femur (316 and 392 N mm-1, respectively) after 8 weeks of implantation. These 3D implants have great potential for patient-specific bone defect treatments.